- Research article
- Open Access
Dimerization of inositol monophosphatase Mycobacterium tuberculosis SuhB is not constitutive, but induced by binding of the activator Mg2+
- Alistair K Brown†1,
- Guoyu Meng†1, 4,
- Hemza Ghadbane†1,
- David J Scott2,
- Lynn G Dover1,
- Jérôme Nigou3,
- Gurdyal S Besra1Email author and
- Klaus Fütterer1Email author
© Brown et al; licensee BioMed Central Ltd. 2007
- Received: 22 June 2007
- Accepted: 28 August 2007
- Published: 28 August 2007
The cell wall of Mycobacterium tuberculosis contains a wide range of phosphatidyl inositol-based glycolipids that play critical structural roles and, in part, govern pathogen-host interactions. Synthesis of phosphatidyl inositol is dependent on free myo-inositol, generated through dephosphorylation of myo-inositol-1-phosphate by inositol monophosphatase (IMPase). Human IMPase, the putative target of lithium therapy, has been studied extensively, but the function of four IMPase-like genes in M. tuberculosis is unclear.
We determined the crystal structure, to 2.6 Å resolution, of the IMPase M. tuberculosis SuhB in the apo form, and analysed self-assembly by analytical ultracentrifugation. Contrary to the paradigm of constitutive dimerization of IMPases, SuhB is predominantly monomeric in the absence of the physiological activator Mg2+, in spite of a conserved fold and apparent dimerization in the crystal. However, Mg2+ concentrations that result in enzymatic activation of SuhB decisively promote dimerization, with the inhibitor Li+ amplifying the effect of Mg2+, but failing to induce dimerization on its own.
The correlation of Mg2+-driven enzymatic activity with dimerization suggests that catalytic activity is linked to the dimer form. Current models of lithium inhibition of IMPases posit that Li+ competes for one of three catalytic Mg2+ sites in the active site, stabilized by a mobile loop at the dimer interface. Our data suggest that Mg2+/Li+-induced ordering of this loop may promote dimerization by expanding the dimer interface of SuhB. The dynamic nature of the monomer-dimer equilibrium may also explain the extended concentration range over which Mg2+ maintains SuhB activity.
- Metal Site
- Dime Interface
- Dime State
- Bury Surface Area
- Inositol Ring
The enzyme inositol monophosphatase (myo-inositol-1-phosphate phosphohydrolase, EC 18.104.22.168, IMPase) has attracted considerable scrutiny since the early 1980s. Then, it was discovered that low millimolar concentrations of Li+ inhibit IMPase-catalysed dephosphorylation of myo-inositol-1-phosphate , which could plausibly explain the marked decrease of myo-inositol in brain tissue following administration of lithium, a veteran therapeutic in manic depression treatment . Inhibition of myo-inositol synthesis affects the phosphatidyl inositol second messenger pathway, which is linked to manic depression, and it is now widely accepted, though unproven, that IMPases constitute a major target of lithium therapy .
The mycobacterial cell wall contains several lipid constituents based on the structure of phosphatidylinositol (PI), such as free PI, phosphatidylinositol-mannosides, lipomannan, and lipoarabinomannan . In addition to their critical structural role, these lipids are significant as immunomodulatory factors in interactions of the tubercle bacillus with the host [5–7]. PI, an essential structural component, is synthesised by phosphatidylinositol synthase, M. tuberculosis PgsA, from CDP-diacylglycerol and myo-inositol . In mycobacteria, the supply of myo-inositol to PI synthesis is thought to be maintained by de novo synthesis, which entails conversion of glucose-6-phosphate to inositol-1-phosphate, catalysed by inositol-1-phosphate synthase, and subsequent dephosphorylation of inositol-1-phosphate catalysed by an IMPase . Thus, IMPase activity is a critical part of PI biosynthesis in mycobacteria. Indeed, growth of Mycobacterium smegmatis is accompanied by IMPase activity that dies off as growth reaches the stationary phase, while growth is retarded in the presence of lithium .
Structure determination of SuhB
ID14-2 (ESRF, Grenoble)
Unit cell parameters
a, b, c (Å)
101.5, 185.4, 106.9
Resolution range (Å)
< I/σ(I) >
No. of non-hydrogen atoms
Average B-factors (Å2)
Main chain – subunit A/B/C
Side-chain – subunit A/B/C
RMSD bonds (Å)
RMSD angles (°)
Comparison of M. tuberculosis SuhB to selected members of the Mg2+-dependent/Li+-inhibited phosphomonoesterase family of proteins
Eukaryotic adenosine-monophosphate (AMP)-regulated FBPases have been described in terms of two sub-domains. In this description, layers I and II of the α-β-α-β-α sandwich are equivalent to the AMP-binding sub-domain of porcine FBPase, and layers III to V correspond to the fructose-1,6-bisphosphate binding sub-domain (Figure 2A) . The primary sequence runs sequentially through layers I and II of the 'AMP-binding' sub-domain, but meanders in the fructose-1,6-bisphosphate-binding sub-domain between layers III-V, with helices and strands alternating. This pattern is interrupted only for strands β12 and β13, which are not separated by a helix (Figures 1, 2A). The dimer observed in the crystal structure of SuhB is primarily mediated by helices α6 in layer V and the β-sheet in layer IV, which extends across the interface (Figure 2B). In contrast, strands in the sheet of layer II are oriented perpendicular to the plane of the dimer interface.
Due to the absence of divalent metal ions and of substrate in the active site, the loop connecting helices α1 and α2 is disordered (residues 35–53 missing), as is much of the loop connecting strands β3 and β4 (residues 85–96 missing). Both features are consistent with the structure of the apo form of human IMPase (pdb entry 1IMF, ). Soaks of SuhB crystals in Mg2+- or Mn2+-containing cryoprotection buffers failed to induce ordering, or give rise to density peaks associated with metals sites known from the eukaryotic orthologues, which we attribute to the acidic pH (4.1–4.5) of the crystallisation buffer. Further disorder was seen in the hairpin loop connecting strands β12 and β13 (residues 254–259) in layer IV, while density for the β9-α6 loop was disjointed, yet indicating that the conformation of the loop between residues 183 and 186 differed markedly between subunits. As a consequence, side chains for residues Tyr183, Val185, Arg186 and Cys187 in this loop could not be built.
The active site of SuhB is situated in a cavity carved out at the N-terminal end of helix α7, sandwiched between layers II and IV (Figure 2A). Due to the disordered α1-α2 loop – referred to as the 'mobile loop'  – the active site in SuhB appears wide open to solvent. Yet, it is anticipated that the α1-α2 loop will become ordered upon the enzyme binding metal ions and substrate. In this state the active site is anticipated to be effectively shielded from solvent, as is the case in the ligand-bound structure of human IMPase . In SuhB, the α1-α2 loop has a 10-residue insertion relative to human IMPase (Figure 1). Hence, the exact conformation of this loop in the ordered state may well deviate from that of the human enzyme. Interestingly, alternative secondary structure conformations of this loop were observed within the tetramer of the structure of Thermotoga maritima IMPase/FBPase TM1415. While the mobile loop included a short helix in two subunits of the tetramer, it assumed a β-hairpin conformation in the other two subunits . The mobile loop includes a lysine residue at position 36 (corresponding to residue 49 in SuhB, marked by I in Figure 1) that stabilizes the third of three catalytic metal sites through hydrogen bonds to two ordered water molecules. In SuhB, this lysine is conserved and parallels in the characteristics of Li+ inhibition between SuhB and human IMPase (see Discussion) suggest an equivalent pattern of interactions in the mycobacterial enzyme.
In our model of the SuhB-substrate complex, the phosphate moiety is positioned at the N-terminal end of helix α4, the helix dipole countering the charge of the phosphate (Figure 3A). Most contacts between enzyme and phosphate are through the three Mg2+ ions, with contact distances in the order of 2.15–2.3 Å, in agreement with the experimental structures of the eukaryotic IMPases [17, 28, 29]. In addition, the phosphate forms H-bond interactions with the amide nitrogen of Gly108 (h_94) and Thr109 (h_95) at the N-terminus of helix α4. The inositol moiety packs primarily against residues in the β9-α6 and β10-α7 loops with specificity-determining contacts provided by Glu228 (h_213) in strand β11, and Asp107 (h_93) in the β4-α4 loop (Figure 3A). All conserved side chains forming the metal binding sites or contacting inositol-1-phosphate are seen in essentially the same conformation as in the human/bovine enzyme, with the exception of Glu83: as a consequence of disorder in a large part of the β3-β4 loop, Glu83 (h_70) in subunit A points outward rather into the active site (Figure 3A).
Significant discrepancies between the M. tuberculosis and the human enzyme are seen in the β9-α6 loop, which in human IMPase provides a contact surface, but no specificity-determining interactions with the inositol ring  (Figure 3B). The β9-α6 loop in SuhB is two residues shorter than in the human enzyme with weak sequence conservation (Figure 1). Although poorly ordered between residues 183–186, residues in proximity to the putative position of the inositol ring – Gly180, Phe181 and Gly182 – are well defined by density (Figure 3B). Interestingly, the carbonyl oxygen of Phe181 falls within H-bonding distance range of the C3 (2.9 Å) and C4 (3.3 Å) hydroxyls of the inositol ring, suggesting that the β9-α6 of SuhB may form specificity-determining contacts to the substrate that are not present in the human enzyme.
A surprising observation was a patch of unexplained density in the active site that overlaps significantly with the modelled position of inositol-1-phosphate, while a second density peak coincides with the metal site 1 (Figure 3C) (in the numbering of ref. ). The latter is situated between the carbonyl oxygen of Ile106 and the carboxylate of Asp104 and corresponds to the high-affinity metal site . The 'metal site 1' peak was present in all three active sites, whereas the density for the inositol-1-phosphate site was best defined in subunit C (Figure 3C). We consider it likely that the observed density represents a weakly-bound molecule of octyl β-D-glucopyranoside, a reagent used as an additive during crystallisation. Whether or not the density peak a metal site 1 represents a divalent cation could not be verified.
Dimer interface and assembly state
The interface of the SuhB dimer buries a total of 2696 Å2 of solvent accessible surface, calculated using the PISA interface server [30, 31], or 1348 Å2 per monomer. In terms of buried surface area hydrophobic interactions clearly dominate over H-bonds (12 contacts). Although the residues located at the interface include a number of polar and charged side chains (Figure 4C) only one ionic interaction is seen. In human IMPase distinctly more solvent accessible surface is buried in the interface (1675 Å2 per monomer). Contacts across the interface include 11 ionic interactions, while the number of H-bonds (13) is about the same as in SuhB. One factor explaining the difference in size of the interface, at least in part, is the disordered 'mobile loop' in SuhB. In the apo structure of human IMPase (1IMF – ), the α1-α2 loop is also disordered, reducing the total of buried solvent accessible surface per monomer from 1675 Å2 to 1571 Å2. Thus disorder of this loop accounts partially for the observed discrepancy.
Next, we examined whether the inhibitor Li+ influenced dimerization. At an enzyme concentration of 1.0 mg.ml-1 and in the presence of 5 mM Mg2+, increasing concentrations of Li+ amplify the effect of Mg2+-induced dimerization, with the dimer peak becoming dominant over the monomer peak (Figure 6B). However on its own Li+ promotes dimerization only very weakly, if at all (Figure 6B). Furthermore the c(S) distribution obtained for 15 mM Mg2+ matches almost perfectly the one obtained in presence of 5 mM Mg2+ and 5 mM Li+. Calcium, which binds to IMPase on identical sites as Mg2+, but does not activate, strongly promotes dimerization, while EDTA reverses Mg2+-induced dimerization [see Additional file 1].
The present crystal structure of M. tuberculosis SuhB confirms a highly conserved structural scaffold of IMPases in mycobacteria with respect to overall fold and active site geometry, in spite of a low level of overall sequence identity (25%) relative to human IMPase. The structural similarity correlates with similar biochemical characteristics – activation by magnesium, inhibition by lithium, specificity for inositol-1-phosphate and exclusion of fructose-1,6-bisphosphate as a substrate. It has been noted previously that the Mg2+-dependence of IMPase activity of SuhB resembles more closely that of the thermophilic species Thermotoga maritima, Archeoglobus fulgidus and Methanococcus jannaschii [12, 25, 33, 34] in that activation is maintained over a concentration range of Mg2+ in the order of 100 mM, whereas in eukaryotic IMPases Mg2+ becomes inhibitory above 5 mM. Such comparison ignores, however, that unlike these thermophilic enzymes SuhB does not hydrolyze fructose-1,6-bisphosphate, and remains sensitive to Li+ (IC50 0.9 mM, ). Given that the active site of SuhB, in its 'non-mobile' part, displays the same highly conserved framework of side chains coordinating the three metal sites as the eukaryotic orthologues, we postulate that differences in Mg2+-dependence of activity between SuhB and eukaryotic IMPases must be linked on the one hand to structural differences in the mobile α1-α2 loop, and secondly to the apparent link between Mg2+-driven dimerization and activation of SuhB. In IMPases and the tetrameric IMPase/FBPases the α1-α2 loop, when ordered, stabilizes the third of the three catalytic metal sites through bridging water molecules [17, 25, 29]. The α1-α2 loop contributes significantly to the dimer interface. This is apparent when comparing the buried solvent-accessible surface in the dimer interface of human IMPase between the ordered and disordered state of the mobile loop. Thus, we anticipate that ordering of the α1-α2 loop in SuhB, which is 10 residues longer compared to human IMPase, will expand the dimer interface and help stabilize the dimer state. While SuhB is driven to the dimer state at high protein concentrations in the absence of metal ions, as was the case during crystallization, it is evident that Mg2+ strongly promotes dimerization at low concentrations. Lithium amplifies the Mg2+-induced effect, yet Li+ alone does not noticeably shift the monomer dimer equilibrium. These observations correlate with the characteristics of metal binding in the active site, which is known to promote ordering of the α1-α2 loop. According to 7Li-NMR binding data, lithium occupies a single site per monomer in IMPase , and various indirect evidence points to Li+ displacing Mg2+ from either site 2 or site 3 [17, 21, 36, 37]. A lysine residue in the mobile loop (Lys36 in human, open asterisk in Figure 1) that coordinates the third metal site via hydrogen bonds to bridging water molecules has been shown to critically influence Li+-sensitivity , and this lysine is conserved in SuhB (Lys49). Given that the α1-α2 loop is disordered, the present structure leaves open the precise geometry of this loop in the ordered state and whether Lys49 in SuhB coordinates the third metal site. Yet, lithium sensitivity of SuhB (IC50 0.9 mM, ), the debilitating effect on activity of the W234A mutation, and the loss of Li+ inhibition through the L81A mutation  – a mutation that removes a stabilising hydrophobic contact to the conserved Trp234 in the active site – hint that metal binding induces ordering of the α1-α2 loop in SuhB in a similar fashion as in the eukaryotic orthologues.
Previously, size exclusion chromatography experiments with 1 mM EDTA present in the buffer had indicated that E. coli SuhB formed monomers and it had been inferred that the E. coli enzyme was active in the monomeric form . The parallels between metal-dependent self-association and enzymatic activation in M. tuberculosis SuhB strongly suggest that dimerization is linked to loading of the three metal sites in the active site, and that phosphatase activity occurs in the dimer state, although we are not in a position to determine whether dimerization is required for activity. If dimerization were required for enzymatic activity of SuhB, this would at least in part explain the wide range of Mg2+ concentrations over which activity is maintained, as the transition to the dimer state is not complete up to at least 15 mM Mg2+. Several caveats go with this rationale. First, the comparison of the activity and AUC data is complicated by the fact that metal binding is cooperative with substrate binding  and inositol-1-phosphate was not present in the AUC experiments. Also, while dimerisation appears to correlate with loading of the metal positions in the active site, we cannot rule out the possibility that the dimer interface contains one or more metal binding sites, which could drive dimerisation. However, the present structure, in line with related IMPases, and a series of metal-soaking experiments with SuhB crystals provided no indication for such a site (data not shown).
Crystal structures of eukaryotic IMPases, a refolding study and AUC analysis of human IMPase 2, in the absence of Mg2+, consistently indicate constitutive dimerization of the eukaryotic orthologue [16, 17, 23, 39]. It is not clear what mechanistic purpose dimerization serves with respect to the enzymatic properties of IMPases, and why dimerization should be constitutive in eukarya, but not in bacteria. Unlike the tetrameric FBPases, which are regulated by an allosteric mechanism involving changes in the relative orientation of the subunits in the tetramer (see  and references therein), no regulatory mechanism for IMPase has been reported that invokes dimerization of the enzyme. Nevertheless, Mg2+ has been shown to moderately enhance thermal stability of M. tuberculosis SuhB, which correlates to some extent with phosphohydrolase activity of SuhB peaking at about 80°C . While this latter property is mirrored by E. coli SuhB  and M. tuberculosis CysQ  it has not been tested whether at such high temperatures SuhB still discriminates between substrates. Thus, while dimerization may increase thermal stability, the functional role of Mg2+-induced dimerization of SuhB in the physiological regime of the tubercle bacillus is not clear.
SuhB – a template for the other M. tuberculosis IMPases?
BLAST search for M. tuberculosis IMPase homologues of known structure
1IMA (IMP human)
2BJI (IMP bovine)
Analysing the multiple sequence alignment that underlies Table 3 [see Additional file 2] more closely, it is interesting to note that enzymes that are specific for inositol-1-phosphate and exclude fructose-1,6-bisphosphate carry a glutamic acid or glutamine at position 228 (SuhB sequence; residue 213 in human IMPase), whereas Asp, His, Ser or Thr are found in enzymes that hydrolyze both substrates or are specific for fructose-1,6-bisphosphate. In the inositol-1-phosphate-bound structure of human IMPase , the corresponding residue, Glu213, forms hydrogen bonds to the C3 and C4 hydroxyls of the inositol ring. In addition to the H-bond between Asp93 (Asp107 in SuhB) and the C6-hydroxyl, these H-bonds are the only side chain-mediated, specificity-determining contacts between the inositol moiety and the enzyme. In the structural superimposition it is evident that an Asp (and likewise Ser, Thr or His) at this position is unlikely to confer selectivity as the distance between the terminal group of the side chain and the sugar hydroxyls becomes too large. Discrimination against fructose-1,6-bisphosphate is rooted primarily in steric clashes of the 6-phosphate the β9-α6 loop. As a result of deletions relative to human IMPase, a much shorter β9-α6 loop in FBPases and dual activity IMPase/FBPases provide the space to accommodate the second phosphate group. Thus, deletions in this region in addition to substitutions of Glu/Gln at position 228 could be indicative of dual specificity. Analysing the sequences of ImpA and Rv3137 in light of these considerations suggests that at least Rv3137 is restricted to inositol-1-phosphate, whereas the case is less clear cut for ImpA. The latter carries a Ser at the position corresponding to Glu228, but displays no significant deletion relative to SuhB between residues 150 and 190, whereas CysQ by the standards of both criteria falls into the category of the dual specificity enzymes, consistent with published biochemical data .
We have determined the structure of M. tuberculosis SuhB, providing a structural template for the four IMPase-like enzymes in this organism. While resembling eukaryotic IMPases in terms of structural scaffold, specificity for inositol-1-phosphate, requirement for Mg2+ and inhibition by Li+, SuhB clearly diverges from the paradigm of constitutive dimerization of bona fide IMPases. The present data support a model of Mg2+-dependent dimerization, where loading of the three catalytic metal sites in the active site induces ordering of the mobile loop, promoting dimerization likely by expanding the dimer interface. The active site of SuhB presents a highly conserved scaffold of side chains forming the metal- and substrate-binding site, yet additional specificity-determining contacts are expected with the weakly conserved β9-α6 loop. Sequence and structural comparisons lead us to predict that the essential gene product Rv3137 represents a bona fide IMPase, while such may or may not be the case for ImpA.
Centricon YM-10 filtration units were obtained from Millipore. E. coli C41(DE3) , acquired from Avidis (France), was used for protein production in this study. Sparse matrix crystallisation screens were obtained from Molecular Dimensions Ltd. All other chemicals were reagent grade or better and obtained from Sigma-Aldrich.
Expression, purification and crystallisation
Recombinant proteins were generated and purified as described previously . Purified His6-tagged SuhB, was dialyzed extensively against 20 mM sodium phosphate pH 7.5, 250 mM NaCl and purified further by size exclusion chromatography over a Sephacryl S200-HR matrix. Pure fractions were pooled and concentrated using Centricon YM-10 filter units. Crystals of SuhB were obtained by hanging drop vapour diffusion, mixing 1 μl of protein solution (40–50 mg.ml-1) with 1 μl of reservoir solution, screening against commercial sparse matrix screens (Structure Screen 1 and 2, Molecular Dimensions Ltd.) at 18°C. Initial crystals appeared in 30%(v/v)2-methyl-2, 4-pentanediol(MPD), 0.1 M tri-sodium citrate, pH 5.6, 0.2 M ammonium acetate. Optimization led to crystals approximately 0.3 mm in length at 10–14%(v/v) MPD, 0.1 M citric acid, pH 4.2, and 0.05%(w/v) octyl β-D-glucopyranoside. Crystals grew to their final size over 7–10 days. In preparation for X-ray data collection crystals were soaked in cryoprotectant , consisting of upto 30%(v/v) MPD, 125 mM NaCl, 0.1 M citric acid pH 4.2 and 0.05%(w/v) octyl β-D-glucopyranoside, increasing the MPD concentration in steps of 10%. Crystals were mounted in nylon loops and frozen in a 100 K nitrogen gas stream.
X-ray data collection and crystallographic analysis
X-ray diffraction data for SuhB were recorded on beamline ID14-EH2 at the European Synchrotron Radiation Facility (ESRF), Grenoble, France (Table 1). Data were reduced using DENZO/SCALEPACK . Patterson self-rotation analysis (POLARRFN – ) revealed a 3-fold non-crystallographic rotation axis parallel to the 21-screw axis (resulting in an apparent 6-fold), consistent with three SuhB monomers per crystallographic asymmetric unit. Structure factor amplitudes were normalized (ECALC – ), and initial phases were obtained by molecular replacement (AMORE – ), using monomer coordinates of human IMPase (PDB:1IMA – ) as a search model. While the Patterson cross-rotation search did not reveal clear solutions, the correct 3 orientations (peaks 4, 5 and 7) could be identified through comparing rotation angles between rotation function peaks with self-rotation peaks, which was confirmed through the subsequent translation search. The search model was stripped of non-conserved side chains and insertions, and built manually into MR-phased density (O – ). Search model bias was minimised by building into simulated annealing omit maps , followed by rounds of refinement (conjugate-gradient minimization, simulated annealing) in CNS ver 1.1  and manual rebuilding, eventually leading to a model that fitted the data with an Rfree of ~30% (5% of reflections). Non-crystallographic symmetry restraints were introduced and the refinement was continued using REFMAC5 . The final model contains 3 monomers covering SuhB residues 5–34, 54–85, 98–253, 260–286. Details of the refinement statistics are listed in Table 1. Coordinates and structure factors have been deposited in the Protein Data Bank  [PDB:2Q74].
Sedimentation velocity experiments were performed using a Beckman Optima XL-A analytical ultracentrifuge equipped with absorbance optics. Protein samples were dialysed into storage buffer, as indicated in the figure legends (Figures 5 and 6) and loaded into cells with two channel Epon centre pieces and quartz windows. Data were recorded at 40,000 rpm, 4°C. A total of 100 absorbance scans (280 nm) were recorded for each sample, representing the full extent of sedimentation of the sample. Data analysis was performed using the SEDFIT software fitting a single friction coefficient .
We thank Tom Burton for his involvement in model building as part of an undergraduate summer project. We thank Tim Dafforn for access to the analytical ultracentrifuge, part of the Biosciences Biomolecular Characterisation Facility (BBCF), and J. Baz Jackson for valuable discussions. We thank ESRF Grenoble for access to the synchrotron and travel support, and ESRF staff for support during data acquisition. G.M. was supported by an Adrian Brown Scholarship. G.S.B. acknowledges support in the form of a Personal Research Chair from Mr. James Bardrick, as a former Lister Institute-Jenner Research Fellow, and the Medical Research Council.
- Hallcher LM, Sherman WR: The effects of lithium ion and other agents on the activity of myo-inositol-1-phosphatase from bovine brain. J Biol Chem 1980, 255: 10896–10901.PubMedGoogle Scholar
- Allison JH, Stewart MA: Reduced brain inositol in lithium-treated rats. Nat New Biol 1971, 233: 267–268. 10.1038/233330a0View ArticlePubMedGoogle Scholar
- Miller DJ, Allemann RK: myo-Inositol monophosphatase: a challenging target for mood stabilising drugs. Mini Rev Med Chem 2007, 7: 107–113. 10.2174/138955707779802624View ArticlePubMedGoogle Scholar
- Brennan PJ, Nikaido H: The envelope of mycobacteria. Annu Rev Biochem 1995, 64: 29–63. 10.1146/annurev.bi.64.070195.000333View ArticlePubMedGoogle Scholar
- Chatterjee D, Khoo KH: Mycobacterial lipoarabinomannan: an extraordinary lipoheteroglycan with profound physiological effects. Glycobiology 1998, 8: 113–120. 10.1093/glycob/8.2.113View ArticlePubMedGoogle Scholar
- Strohmeier GR, Fenton MJ: Roles of lipoarabinomannan in the pathogenesis of tuberculosis. Microbes Infect 1999, 1: 709–717. 10.1016/S1286-4579(99)80072-0View ArticlePubMedGoogle Scholar
- Vercellone A, Nigou J, Puzo G: Relationships between the structure and the roles of lipoarabinomannans and related glycoconjugates in tuberculosis pathogenesis. Front Biosci 1998, 3: e149–63.PubMedGoogle Scholar
- Jackson M, Crick DC, Brennan PJ: Phosphatidylinositol is an essential phospholipid of mycobacteria. J Biol Chem 2000, 275: 30092–30099. 10.1074/jbc.M004658200View ArticlePubMedGoogle Scholar
- Chen IW, Charalampous CF: Biochemical studies on inositol. IX. D-Inositol 1-phosphate as intermediate in the biosynthesis of inositol from glucose 6-phosphate, and characteristics of two reactions in this biosynthesis. J Biol Chem 1966, 241: 2194–2199.PubMedGoogle Scholar
- Nigou J, Besra GS: Characterization and regulation of inositol monophosphatase activity in Mycobacterium smegmatis. Biochem J 2002, 361: 385–390.PubMed CentralView ArticlePubMedGoogle Scholar
- Cole ST, Brosch R, Parkhill J, Garnier T, Churcher C, Harris D, Gordon SV, Eiglmeier K, Gas S, Barry CEr, Tekaia F, Badcock K, Basham D, Brown D, Chillingworth T, Connor R, Davies R, Devlin K, Feltwell T, Gentles S, Hamlin N, Holroyd S, Hornsby T, Jagels K, Krogh A, McLean J, Moule S, Murphy L, Oliver K, Osborne J, Quail MA, Rajandream MA, Rogers J, Rutter S, Seeger K, Skelton J, Squares R, Squares S, Sulston JE, Taylor K, Whitehead S, Barrell BG: Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature 1998, 393: 537–544. 10.1038/31159View ArticlePubMedGoogle Scholar
- Nigou J, Dover LG, Besra GS: Purification and biochemical characterization of Mycobacterium tuberculosis SuhB, an inositol monophosphatase involved in inositol biosynthesis. Biochemistry 2002, 41: 4392–4398. 10.1021/bi0160056View ArticlePubMedGoogle Scholar
- Chen L, Roberts MF: Overexpression, purification, and analysis of complementation behavior of E. coli SuhB protein: comparison with bacterial and archaeal inositol monophosphatases. Biochemistry 2000, 39: 4145–4153. 10.1021/bi992424fView ArticlePubMedGoogle Scholar
- Sassetti CM, Boyd DH, Rubin EJ: Genes required for mycobacterial growth defined by high density mutagenesis. Mol Microbiol 2003, 48: 77–84. 10.1046/j.1365-2958.2003.03425.xView ArticlePubMedGoogle Scholar
- Gu X, Chen M, Shen H, Jiang X, Huang Y, Wang H: Rv2131c gene product: an unconventional enzyme that is both inositol monophosphatase and fructose-1,6-bisphosphatase. Biochem Biophys Res Commun 2006, 339: 897–904. 10.1016/j.bbrc.2005.11.088View ArticlePubMedGoogle Scholar
- Bone R, Springer JP, Atack JR: Structure of inositol monophosphatase, the putative target of lithium therapy. Proc Natl Acad Sci U S A 1992, 89: 10031–10035. 10.1073/pnas.89.21.10031PubMed CentralView ArticlePubMedGoogle Scholar
- Gill R, Mohammed F, Badyal R, Coates L, Erskine P, Thompson D, Cooper J, Gore M, Wood S: High-resolution structure of myo-inositol monophosphatase, the putative target of lithium therapy. Acta Crystallogr D Biol Crystallogr 2005, 61: 545–555. 10.1107/S0907444905004038View ArticlePubMedGoogle Scholar
- Stec B, Yang H, Johnson KA, Chen L, Roberts MF: MJ0109 is an enzyme that is both an inositol monophosphatase and the 'missing' archaeal fructose-1,6-bisphosphatase. Nat Struct Biol 2000, 7: 1046–1050. 10.1038/80968View ArticlePubMedGoogle Scholar
- York JD, Ponder JW, Majerus PW: Definition of a metal-dependent/Li(+)-inhibited phosphomonoesterase protein family based upon a conserved three-dimensional core structure. Proc Natl Acad Sci U S A 1995, 92: 5149–5153. 10.1073/pnas.92.11.5149PubMed CentralView ArticlePubMedGoogle Scholar
- Zhang Y, Liang JY, Lipscomb WN: Structural similarities between fructose-1,6-bisphosphatase and inositol monophosphatase. Biochem Biophys Res Commun 1993, 190: 1080–1083. 10.1006/bbrc.1993.1159View ArticlePubMedGoogle Scholar
- Albert A, Yenush L, Gil-Mascarell MR, Rodriguez PL, Patel S, Martinez-Ripoll M, Blundell TL, Serrano R: X-ray structure of yeast Hal2p, a major target of lithium and sodium toxicity, and identification of framework interactions determining cation sensitivity. J Mol Biol 2000, 295: 927–938. 10.1006/jmbi.1999.3408View ArticlePubMedGoogle Scholar
- Patel S, Yenush L, Rodriguez PL, Serrano R, Blundell TL: Crystal structure of an enzyme displaying both inositol-polyphosphate-1-phosphatase and 3'-phosphoadenosine-5'-phosphate phosphatase activities: a novel target of lithium therapy. J Mol Biol 2002, 315: 677–685. 10.1006/jmbi.2001.5271View ArticlePubMedGoogle Scholar
- Arai R, Ito K, Ohnishi T, Ohba H, Akasaka R, Bessho Y, Hanawa-Suetsugu K, Yoshikawa T, Shirouzu M, Yokoyama S: Crystal structure of human myo-inositol monophosphatase 2, the product of the putative susceptibility gene for bipolar disorder, schizophrenia, and febrile seizures. Proteins 2007, 67: 732–742. 10.1002/prot.21299View ArticlePubMedGoogle Scholar
- Johnson KA, Chen L, Yang H, Roberts MF, Stec B: Crystal structure and catalytic mechanism of the MJ0109 gene product: a bifunctional enzyme with inositol monophosphatase and fructose 1,6-bisphosphatase activities. Biochemistry 2001, 40: 618–630. 10.1021/bi0016422View ArticlePubMedGoogle Scholar
- Stieglitz KA, Johnson KA, Yang H, Roberts MF, Seaton BA, Head JF, Stec B: Crystal structure of a dual activity IMPase/FBPase (AF2372) from Archaeoglobus fulgidus. The story of a mobile loop. J Biol Chem 2002, 277: 22863–22874. 10.1074/jbc.M201042200View ArticlePubMedGoogle Scholar
- Ke HM, Zhang YP, Lipscomb WN: Crystal structure of fructose-1,6-bisphosphatase complexed with fructose 6-phosphate, AMP, and magnesium. Proc Natl Acad Sci U S A 1990, 87: 5243–5247. 10.1073/pnas.87.14.5243PubMed CentralView ArticlePubMedGoogle Scholar
- Stieglitz KA, Roberts MF, Li W, Stec B: Crystal structure of the tetrameric inositol 1-phosphate phosphatase (TM1415) from the hyperthermophile, Thermotoga maritima. FEBS J 2007, 274: 2461–2469. 10.1111/j.0014-2956.2007.05779.xView ArticlePubMedGoogle Scholar
- Bone R, Frank L, Springer JP, Atack JR: Structural studies of metal binding by inositol monophosphatase: evidence for two-metal ion catalysis. Biochemistry 1994, 33: 9468–9476. 10.1021/bi00198a012View ArticlePubMedGoogle Scholar
- Bone R, Frank L, Springer JP, Pollack SJ, Osborne SA, Atack JR, Knowles MR, McAllister G, Ragan CI, Broughton HB, et al.: Structural analysis of inositol monophosphatase complexes with substrates. Biochemistry 1994, 33: 9460–9467. 10.1021/bi00198a011View ArticlePubMedGoogle Scholar
- Protein interfaces, surfaces and assemblies service PISA at European Bioinformatics Institute[http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html]
- Krissinel E, Henrick K: Inference of macromolecular assemblies from crystallline state. J Mol Biol 2007. doi:10.1016/j.jmb.2007.05.022:available online 13 May 2007.Google Scholar
- Schuck P: On the analysis of protein self-association by sedimentation velocity analytical ultracentrifugation. Anal Biochem 2003, 320: 104–124. 10.1016/S0003-2697(03)00289-6View ArticlePubMedGoogle Scholar
- Chen L, Roberts MF: Cloning and expression of the inositol monophosphatase gene from Methanococcus jannaschii and characterization of the enzyme. Appl Environ Microbiol 1998, 64: 2609–2615.PubMed CentralPubMedGoogle Scholar
- Chen L, Roberts MF: Characterization of a tetrameric inositol monophosphatase from the hyperthermophilic bacterium Thermotoga maritima. Appl Environ Microbiol 1999, 65: 4559–4567.PubMed CentralPubMedGoogle Scholar
- Saudek V, Vicendon P, Do QT, Atkinson RA, Sklenar V, Pelton PD, Piriou F, Ganzhorn AJ: 7Li nuclear-magnetic-resonance study of lithium binding to myo-inositolmonophosphatase. Eur J Biochem 1996, 240: 288–291.View ArticlePubMedGoogle Scholar
- Ganzhorn AJ, Chanal MC: Kinetic studies with myo-inositol monophosphatase from bovine brain. Biochemistry 1990, 29: 6065–6071. 10.1021/bi00477a026View ArticlePubMedGoogle Scholar
- Ganzhorn AJ, Lepage P, Pelton PD, Strasser F, Vincendon P, Rondeau JM: The contribution of lysine-36 to catalysis by human myo-inositol monophosphatase. Biochemistry 1996, 35: 10957–10966. 10.1021/bi9603837View ArticlePubMedGoogle Scholar
- Strasser F, Pelton PD, Ganzhorn AJ: Kinetic characterization of enzyme forms involved in metal ion activation and inhibition of myo-inositol monophosphatase. Biochem J 1995, 307: 585–593.PubMed CentralView ArticlePubMedGoogle Scholar
- Kwon OS, Lo SC, Kwok F, Churchich JE: Reversible unfolding of myo-inositol monophosphatase. J Biol Chem 1993, 268: 7912–7916.PubMedGoogle Scholar
- Hines JK, Fromm HJ, Honzatko RB: Novel allosteric activation site in Escherichia coli fructose-1,6-bisphosphatase. J Biol Chem 2006, 281: 18386–18393. 10.1074/jbc.M602553200View ArticlePubMedGoogle Scholar
- Miroux B, Walker JE: Over-production of proteins in Escherichia coli: mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels. J Mol Biol 1996, 260: 289–298. 10.1006/jmbi.1996.0399View ArticlePubMedGoogle Scholar
- Otwinowski Z, Minor W: Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol 1997, 276: 307–326.View ArticleGoogle Scholar
- CCP4: The CCP4 suite: programs for protein crystallography. Acta Crystallogr D Biol Crystallogr 1994, 50: 760–763. 10.1107/S0907444994003112View ArticleGoogle Scholar
- Jones TA, Zou JY, Cowan SW, Kjeldgaard M: Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr A 1991, 47: 110–119. 10.1107/S0108767390010224View ArticlePubMedGoogle Scholar
- Hodel A, Kim S-H, Brünger AT: Model bias in crystal structures. Acta Crystallogr A 1992, 48: 851–858. 10.1107/S0108767392006044View ArticleGoogle Scholar
- Brunger AT, Adams PD, Clore GM, DeLano WL, Gros P, Grosse-Kunstleve RW, Jiang JS, Kuszewski J, Nilges M, Pannu NS, Read RJ, Rice LM, Simonson T, Warren GL: Crystallography & NMR system: A new software suite for macromolecular structure determination. Acta Crystallogr D Biol Crystallogr 1998, 54: 905–921. 10.1107/S0907444998003254View ArticlePubMedGoogle Scholar
- Murshudov GN, Vagin AA, Dodson EJ: Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallogr D Biol Crystallogr 1997, 53: 240–255. 10.1107/S0907444996012255View ArticlePubMedGoogle Scholar
- The RCSB Protein Data Bank (PDB)[http://www.rcsb.org]
- The PyMOL Molecular Graphics System[http://www.pymol.org]
- Schwede T, Diemand A, Guex N, Peitsch MC: Protein structure computing in the genomic era. Res Microbiol 2000, 151: 107–112. 10.1016/S0923-2508(00)00121-2View ArticlePubMedGoogle Scholar
- PovRay – Persistence of Vision Raytracer[http://www.povray.org]
- Schuck P: Size-distribution analysis of macromolecules by sedimentation velocity ultracentrifugation and lamm equation modeling. Biophys J 2000, 78: 1606–1619.PubMed CentralView ArticlePubMedGoogle Scholar
- Gille C, Frommel C: STRAP: editor for STRuctural Alignments of Proteins. Bioinformatics 2001, 17: 377–378. 10.1093/bioinformatics/17.4.377View ArticlePubMedGoogle Scholar
- ESPript, Easy Sequencing in Postscript[http://espript.ibcp.fr/ESPript/ESPript/]
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