1lg5 Citations

Organization of an efficient carbonic anhydrase: implications for the mechanism based on structure-function studies of a T199P/C206S mutant.

Biochemistry 41 7628-35 (2002)
Related entries: 1lg6, 1lgd

Cited: 14 times
EuropePMC logo PMID: 12056894

Abstract

Substitution of Pro for Thr199 in the active site of human carbonic anhydrase II (HCA II)(1) reduces its catalytic efficiency about 3000-fold. X-ray crystallographic structures of the T199P/C206S variant have been determined in complex with the substrate bicarbonate and with the inhibitors thiocyanate and beta-mercaptoethanol. The latter molecule is normally not an inhibitor of wild-type HCA II. All three ligands display novel binding interactions to the T199P/C206S mutant. The beta-mercaptoethanol molecule binds in the active site area with its sulfur atom tetrahedrally coordinated to the zinc ion. Thiocyanate binds tetrahedrally coordinated to the zinc ion in T199P/C206S, in contrast to its pentacoordinated binding to the zinc ion in wild-type HCA II. Bicarbonate binds to the mutant with two of its oxygens at the positions of the zinc water (Wat263) and Wat318 in wild-type HCA II. The environment of this area is more hydrophilic than the normal bicarbonate-binding site of HCA II situated in the hydrophobic part of the cavity normally occupied by the so-called deep water (Wat338). The observation of a new binding site for bicarbonate has implications for understanding the mechanism by which the main-chain amino group of Thr199 acquired an important role for orientation of the substrate during the evolution of the enzyme.

Articles - 1lg5 mentioned but not cited (4)

  1. Direct binding of the EGF-like domain of neuregulin-1 to integrins ({alpha}v{beta}3 and {alpha}6{beta}4) is involved in neuregulin-1/ErbB signaling. Ieguchi K, Fujita M, Ma Z, Davari P, Taniguchi Y, Sekiguchi K, Wang B, Takada YK, Takada Y. J Biol Chem 285 31388-31398 (2010)
  2. Integrins αvβ3 and α4β1 act as coreceptors for fractalkine, and the integrin-binding defective mutant of fractalkine is an antagonist of CX3CR1. Fujita M, Takada YK, Takada Y. J Immunol 189 5809-5819 (2012)
  3. The integrin-binding defective FGF2 mutants potently suppress FGF2 signalling and angiogenesis. Mori S, Hatori N, Kawaguchi N, Hamada Y, Shih TC, Wu CY, Lam KS, Matsuura N, Yamamoto H, Takada YK, Takada Y. Biosci Rep 37 BSR20170173 (2017)
  4. The binding of monomeric C-reactive protein (mCRP) to Integrins αvβ3 and α4β1 is related to its pro-inflammatory action. Fujita M, Takada YK, Izumiya Y, Takada Y. PLoS One 9 e93738 (2014)


Reviews citing this publication (2)

  1. Carbonic anhydrase as a model for biophysical and physical-organic studies of proteins and protein-ligand binding. Krishnamurthy VM, Kaufman GK, Urbach AR, Gitlin I, Gudiksen KL, Weibel DB, Whitesides GM. Chem Rev 108 946-1051 (2008)
  2. Thermodynamic, kinetic, and structural parameterization of human carbonic anhydrase interactions toward enhanced inhibitor design. Linkuvienė V, Zubrienė A, Manakova E, Petrauskas V, Baranauskienė L, Zakšauskas A, Smirnov A, Gražulis S, Ladbury JE, Matulis D. Q Rev Biophys 51 e10 (2018)

Articles citing this publication (8)

  1. Structural study of X-ray induced activation of carbonic anhydrase. Sjöblom B, Polentarutti M, Djinovic-Carugo K. Proc Natl Acad Sci U S A 106 10609-10613 (2009)
  2. Kinetic and structural characterization of thermostabilized mutants of human carbonic anhydrase II. Fisher Z, Boone CD, Biswas SM, Venkatakrishnan B, Aggarwal M, Tu C, Agbandje-McKenna M, Silverman D, McKenna R. Protein Eng Des Sel 25 347-355 (2012)
  3. X-ray structure and mutational analysis of the atrazine Chlorohydrolase TrzN. Seffernick JL, Reynolds E, Fedorov AA, Fedorov E, Almo SC, Sadowsky MJ, Wackett LP. J Biol Chem 285 30606-30614 (2010)
  4. The Crystal Structure of a hCA VII Variant Provides Insights into the Molecular Determinants Responsible for Its Catalytic Behavior. Buonanno M, Di Fiore A, Langella E, D'Ambrosio K, Supuran CT, Monti SM, De Simone G. Int J Mol Sci 19 E1571 (2018)
  5. Structural and kinetic effects on changes in the CO(2) binding pocket of human carbonic anhydrase II. West D, Kim CU, Tu C, Robbins AH, Gruner SM, Silverman DN, McKenna R. Biochemistry 51 9156-9163 (2012)
  6. The Loss and Gain of Functional Amino Acid Residues Is a Common Mechanism Causing Human Inherited Disease. Lugo-Martinez J, Pejaver V, Pagel KA, Jain S, Mort M, Cooper DN, Mooney SD, Radivojac P. PLoS Comput Biol 12 e1005091 (2016)
  7. Exploration of the residues modulating the catalytic features of human carbonic anhydrase XIII by a site-specific mutagenesis approach. De Simone G, Di Fiore A, Truppo E, Langella E, Vullo D, Supuran CT, Monti SM. J Enzyme Inhib Med Chem 34 1506-1510 (2019)
  8. Structural insights into the effect of active-site mutation on the catalytic mechanism of carbonic anhydrase. Kim JK, Lee C, Lim SW, Andring JT, Adhikari A, McKenna R, Kim CU. IUCrJ 7 985-994 (2020)