1bwp Citations

Probing the substrate specificity of the intracellular brain platelet-activating factor acetylhydrolase.

Protein Eng. 12 693-700 (1999)
Related entries: 1bwq, 1bwr

Cited: 16 times
EuropePMC logo PMID: 10469831

Abstract

Platelet-activating factor acetylhydrolases (PAF-AHs) are unique PLA2s which hydrolyze the sn-2 ester linkage in PAF-like phospholipids with a marked preference for very short acyl chains, typically acetyl. The recent solution of the crystal structure of the alpha(1) catalytic subunit of isoform Ib of bovine brain intracellular PAF-AH at 1.7 A resolution paved the way for a detailed examination of the molecular basis of substrate specificity in this enzyme. The crystal structure suggests that the side chains of Thr103, Leu48 and Leu194 are involved in substrate recognition. Three single site mutants (L48A, T103S and L194A) were overexpressed and their structures were solved to 2.3 A resolution or better by X-ray diffraction methods. Enzyme kinetics showed that, compared with wild-type protein, all three mutants have higher relative activity against phospholipids with sn-2 acyl chains longer than an acetyl. However, for each of the mutants we observed an unexpected and substantial reduction in the V(max) of the reaction. These results are consistent with the model in which residues Leu48, Thr103 and Leu194 indeed contribute to substrate specificity and in addition suggest that the integrity of the specificity pocket is critical for the expression of full catalytic function, thus conferring very high substrate selectivity on the enzyme.

Reviews citing this publication (3)

  1. A PLA1-2 punch regulates the Golgi complex. Bechler ME, de Figueiredo P, Brown WJ. Trends Cell Biol. 22 116-124 (2012)
  2. Phospholipase A2 enzymes: physical structure, biological function, disease implication, chemical inhibition, and therapeutic intervention. Dennis EA, Cao J, Hsu YH, Magrioti V, Kokotos G. Chem. Rev. 111 6130-6185 (2011)
  3. GDSL family of serine esterases/lipases. Akoh CC, Lee GC, Liaw YC, Huang TH, Shaw JF. Prog. Lipid Res. 43 534-552 (2004)

Articles citing this publication (13)

  1. Rhamnogalacturonan acetylesterase elucidates the structure and function of a new family of hydrolases. Mølgaard A, Kauppinen S, Larsen S. Structure 8 373-383 (2000)
  2. Convergent evolution of enzyme active sites is not a rare phenomenon. Gherardini PF, Wass MN, Helmer-Citterich M, Sternberg MJ. J. Mol. Biol. 372 817-845 (2007)
  3. The emerging roles of PAF acetylhydrolase. McIntyre TM, Prescott SM, Stafforini DM. J. Lipid Res. 50 Suppl S255-9 (2009)
  4. Crystal structure of Escherichia coli thioesterase I/protease I/lysophospholipase L1: consensus sequence blocks constitute the catalytic center of SGNH-hydrolases through a conserved hydrogen bond network. Lo YC, Lin SC, Shaw JF, Liaw YC. J. Mol. Biol. 330 539-551 (2003)
  5. Quantitative comparison of catalytic mechanisms and overall reactions in convergently evolved enzymes: implications for classification of enzyme function. Almonacid DE, Yera ER, Mitchell JB, Babbitt PC. PLoS Comput. Biol. 6 e1000700 (2010)
  6. Using reaction mechanism to measure enzyme similarity. O'Boyle NM, Holliday GL, Almonacid DE, Mitchell JB. J. Mol. Biol. 368 1484-1499 (2007)
  7. Rapid evolution in conformational space: a study of loop regions in a ubiquitous GTP binding domain. Blouin C, Butt D, Roger AJ. Protein Sci. 13 608-616 (2004)
  8. Intracellular erythrocyte platelet-activating factor acetylhydrolase I inactivates aspirin in blood. Zhou G, Marathe GK, Willard B, McIntyre TM. J. Biol. Chem. 286 34820-34829 (2011)
  9. YesT: a new rhamnogalacturonan acetyl esterase from Bacillus subtilis. Martínez-Martínez I, Navarro-Fernández J, Daniel Lozada-Ramírez J, García-Carmona F, Sánchez-Ferrer A. Proteins 71 379-388 (2008)
  10. Crystal structure of yeast YHR049W/FSH1, a member of the serine hydrolase family. Quevillon-Cheruel S, Leulliot N, Graille M, Hervouet N, Coste F, Bénédetti H, Zelwer C, Janin J, Van Tilbeurgh H. Protein Sci. 14 1350-1356 (2005)
  11. A measure of the broad substrate specificity of enzymes based on 'duplicate' catalytic residues. Chakraborty S, Ásgeirsson B, Rao BJ. PLoS ONE 7 e49313 (2012)
  12. Annotation of proteins of unknown function: initial enzyme results. McKay T, Hart K, Horn A, Kessler H, Dodge G, Bardhi K, Bardhi K, Mills JL, Bernstein HJ, Craig PA. J. Struct. Funct. Genomics 16 43-54 (2015)
  13. Evolutionary and structural analyses of heterodimeric proteins composed of subunits with same fold. Sudha G, Naveenkumar N, Srinivasan N. Proteins 83 1766-1786 (2015)


Related citations provided by authors (1)

  1. Brain Acetylhydrolase that Inactivates Platelet-Activating Factor is a G-Protein-Like Trimer. Ho YS, Swenson L, Derewenda U, Serre L, Wei Y, Dauter Z, Hattori M, Adachi T, Aoki J, Arai H, Inoue K, Derewenda ZS Nature 385 89- (1997)