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NMR derived solution structure of an EF-hand calcium-binding protein from Entamoeba Histolytica.

Atreya HS, Sahu SC, Bhattacharya A, Chary KV, Govil G
Biochemistry 40 14392-403 (2001)
Cited: 22 times
EuropePMC logo PMID: 11724551

Abstract

We present the three-dimensional (3D) solution structure of a calcium-binding protein from Entamoeba histolytica (EhCaBP), an etiologic agent of amoebiasis affecting millions worldwide. EhCaBP is a 14.7 kDa (134 residues) monomeric protein thought to play a role in the pathogenesis of amoebiasis. The 3D structure of Ca(2+)-bound EhCaBP has been derived using multidimensional nuclear magnetic resonance (NMR) spectroscopic techniques. The study reveals the presence of two globular domains connected by a flexible linker region spanning 8 amino acid residues. Each domain consists of a pair of helix-loop-helix motifs similar to the canonical EF-hand motif of calcium-binding proteins. EhCaBP binds to four Ca(2+) with high affinity (two in each domain), and it is structurally related to calmodulin (CaM) and troponin C (TnC) despite its low sequence homology ( approximately 29%) with these proteins. NMR-derived structures of EhCaBP converge within each domain with low RMSDs and angular order-parameters for backbone torsion angles close to 1.0. However, the presence of a highly flexible central linker region results in an ill-defined orientation of the two domains relative to one other. These findings are supported by backbone (15)N relaxation rate measurements and deuterium exchange studies, which reveal low structural order parameters for residues in the central linker region. Earlier, biochemical studies showed that EhCaBP is involved in a novel signal transduction mechanism, distinct from CaM. A possible reason for such a functional diversity is revealed by a detailed comparison of the 3D structure of EhCaBP with that of CaM and TnC. The studies indicate a more open C-terminal domain for EhCaBP with larger water exposed total hydrophobic surface area as compared to CaM and TnC. Further dissimilarities between the structures include the presence of two Gly residues (G63 and G67) in the central linker region of EhCaBP, which seem to impart it a greater flexibility compared to CaM and TnC and also play crucial role in its biological function. Thus, unlike in CaM and TnC, wherein the length and/or composition of the central linker have been found to be crucial for their function, in EhCaBP, both flexibility as well as amino acid composition is required for the function of the protein.

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Reviews citing this publication (3)

  1. Calcium-binding proteins of Entamoeba histolytica. Bhattacharya A, Padhan N, Jain R, Bhattacharya S. Arch Med Res 37 221-225 (2006)
  2. Identification of C-terminal neighbours of amino acid residues without an aliphatic 13Cgamma as an aid to NMR assignments in proteins. Barnwal RP, Rout AK, Atreya HS, Chary KV. J Biomol NMR 41 191-197 (2008)
  3. Structural and functional diversity of Entamoeba histolytica calcium-binding proteins. Kumar S, Mishra S, Gourinath S. Biophys Rev (2020)

Articles citing this publication (18)

  1. Calcium-binding protein 1 of Entamoeba histolytica transiently associates with phagocytic cups in a calcium-independent manner. Jain R, Santi-Rocca J, Padhan N, Bhattacharya S, Guillen N, Bhattacharya A. Cell Microbiol 10 1373-1389 (2008)
  2. Structure and inactivation of triosephosphate isomerase from Entamoeba histolytica. Rodríguez-Romero A, Hernández-Santoyo A, del Pozo Yauner L, Kornhauser A, Fernández-Velasco DA. J Mol Biol 322 669-675 (2002)
  3. The C-terminal tail of human neuronal calcium sensor 1 regulates the conformational stability of the Ca²⁺₋ activated state. Heidarsson PO, Bjerrum-Bohr IJ, Jensen GA, Pongs O, Finn BE, Poulsen FM, Kragelund BB. J Mol Biol 417 51-64 (2012)
  4. Rapid measurement of 3J(H N-H alpha) and 3J(N-H beta) coupling constants in polypeptides. Barnwal RP, Rout AK, Chary KV, Atreya HS. J Biomol NMR 39 259-263 (2007)
  5. Crystal structure of calcium binding protein-1 from Entamoeba histolytica: a novel arrangement of EF hand motifs. Kumar S, Padhan N, Alam N, Gourinath S. Proteins 68 990-998 (2007)
  6. Metal binding properties of fluorescent analogues of trichogin GA IV: a conformational study by time-resolved spectroscopy and molecular mechanics investigations. Venanzi M, Bocchinfuso G, Gatto E, Palleschi A, Stella L, Formaggio F, Toniolo C. Chembiochem 10 91-97 (2009)
  7. Chemical shift based editing of CH3 groups in fractionally 13C-labelled proteins using GFT (3, 2)D CT-HCCH-COSY: stereospecific assignments of CH3 groups of Val and Leu residues. Barnwal RP, Atreya HS, Chary KV. J Biomol NMR 42 149-154 (2008)
  8. Crystal structure and trimer-monomer transition of N-terminal domain of EhCaBP1 from Entamoeba histolytica. Kumar S, Ahmad E, Mansuri MS, Kumar S, Jain R, Khan RH, Gourinath S. Biophys J 98 2933-2942 (2010)
  9. Structure of Ca2+-binding protein-6 from Entamoeba histolytica and its involvement in trophozoite proliferation regulation. Verma D, Murmu A, Gourinath S, Bhattacharya A, Chary KVR. PLoS Pathog 13 e1006332 (2017)
  10. Differential native state ruggedness of the two Ca2+-binding domains in a Ca2+ sensor protein. Mohan PM, Mukherjee S, Chary KV. Proteins 70 1147-1153 (2008)
  11. Structural characterization of a novel Ca2+-binding protein from Entamoeba histolytica: structural basis for the observed functional differences with its isoform. Mustafi SM, Mutalik RB, Jain R, Chandra K, Bhattacharya A, Chary KV. J Biol Inorg Chem 14 471-483 (2009)
  12. Structure prediction of a multi-domain EF-hand Ca2+ binding protein by PROPAINOR. Jyothi S, Mustafi SM, Chary KV, Joshi RR. J Mol Model 11 481-488 (2005)
  13. Functional manipulation of a calcium-binding protein from Entamoeba histolytica guided by paramagnetic NMR. Rout AK, Patel S, Somlata, Shukla M, Saraswathi D, Bhattacharya A, Chary KV. J Biol Chem 288 23473-23487 (2013)
  14. Structural basis for the observed differential magnetic anisotropic tensorial values in calcium binding proteins. Mustafi SM, Mukherjee S, Chary KV, Cavallaro G. Proteins 65 656-669 (2006)
  15. Root-mean-square-deviation-based rapid backbone resonance assignments in proteins. Rout AK, Barnwal RP, Agarwal G, Chary KV. Magn Reson Chem 48 793-797 (2010)
  16. Biophysical Reviews' "Meet the Councilor Series"-a profile of Kandala V. R. Chary. Chary KVR. Biophys Rev 13 821-824 (2021)
  17. Letter Sequence specific 1HN, 13C and 15N resonance assignments of a novel calcium-binding protein from Entamoeba histolytica. Bansal-Mutalik R, Mustafi SM, Bhattacharya A, Chary KV. J Biomol NMR 31 379-380 (2005)
  18. Site-specific free energy changes in proteins upon ligand binding by nuclear magnetic resonance: Ca2+ -displacement by Ln3+ in a Ca2+ -binding protein from Entamoeba histolytica. Chandra K, Mustafi SM, Muthukumar S, Chary KV. Chem Biol Drug Des 77 272-280 (2011)


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