PDBsum entry: 1bvm

Hydrolase

PDB id: 1bvm

Contents

Protein chain

123 a.a. *

* Residue conservation analysis

PDB id:

1bvm

Name:

Hydrolase

Title:

Solution nmr structure of bovine pancreatic phospholipase a2, 20 structures

Structure:

Protein (phospholipase a2). Chain: a. Engineered: yes

Source:

Bos taurus. Cattle. Organism_taxid: 9913. Expressed in: escherichia coli. Expression_system_taxid: 562. Other_details: bovine pancreas

NMR struc:

20 models

Authors:

C.-H.Yuan,I.-J.L.Byeon,Y.Li,M.-D.Tsai

Key ref:

C.Yuan et al. (1999). Structural analysis of phospholipase A2 from functional perspective. 1. Functionally relevant solution structure and roles of the hydrogen-bonding network. Biochemistry, 38, 2909-2918. PubMed id: 10074343 DOI: 10.1021/bi982211a

Date:

14-Sep-98 Release date: 16-Sep-99

Protein chain

P00593  (PA21B_BOVIN) -  Phospholipase A2

Seq: 145 a.a.

Struc: 123 a.a.

Enzyme reactions

• Enzyme class: E.C.3.1.1.4 - Phospholipase A(2).
• Reaction: Phosphatidylcholine + H₂O = 1-acylglycerophosphocholine + a carboxylate
• Cofactor: Calcium

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

 Gene Ontology (GO) functional annotation 

• Cellular component: extracellular region (2 terms)
• Biological process: metabolic process (7 terms)
• Biochemical function: hydrolase activity (7 terms)

reference

DOI no: 10.1021/bi982211a

Biochemistry 38:2909-2918 (1999)

PubMed id: 10074343

Structural analysis of phospholipase A2 from functional perspective. 1. Functionally relevant solution structure and roles of the hydrogen-bonding network.

C.Yuan, I.J.Byeon, Y.Li, M.D.Tsai.

ABSTRACT

Bovine pancreatic phospholipase A2 (PLA2), a small (13.8 kDa) Ca2+-dependent lipolytic enzyme, is rich in functional and structural character. In an effort to examine its detailed structure-function relationship, we determined its solution structure by multidimensional nuclear magnetic resonance (NMR) spectroscopy at a functionally relevant pH. An ensemble of 20 structures generated has an average root-mean-square deviation (RMSD) of 0.62 +/- 0.08 A for backbone (N, Calpha, C) atoms and 0.98 +/- 0.09 A for all heavy atoms. The overall structure shows several notable differences from the crystal structure: the first three residues at the N-terminus, the calcium-binding loop (Y25-T36), and the surface loop (V63-N72) appear to be flexible; the alpha-helical conformation of helix B (E17-F22) is absent; helix D appears to be shorter (D59-V63 instead of D59-D66); and the hydrogen-bonding network is less defined. These differences were analyzed in relation to the function of PLA2. We then further examined the H-bonding network, because its functional role or even its existence in solution has been in dispute recently. Our results show that part of the H-bonding network (the portion away from N-terminus) clearly exists in solution, as evidenced by direct observation (at 11.1 ppm) of a strong H-bond between Y73 and D99 and an implicated interaction between D99 and H48. Analyses of a series of mutants indicated that the existence of the Y73.D99 H-bond correlates directly with the conformational stability of the mutant. Loss of this H-bond results in a loss of 2-3 kcal/mol in the conformational stability of PLA2. The unequivocal identification and demonstration of the structural importance of a specific hydrogen bond, and the magnitude of its contribution to conformational stability, are uncommon to the best of our knowledge. Our results also suggest that, while the D99.H48 catalytic diad is the key catalytic machinery of PLA2, it also helps to maintain conformational integrity.