Changes in activity of porcine phospholipase A2 brought about by charge engineering of a major structural element to alter stability.


Abstract

We have modified the stability of porcine phospholipase A2 by charge engineering. The mutations are situated at the N-terminal of a major helix and are N89D and N89D/E92Q. This engineering has significantly altered the activity of the enzyme to aggregated and monomeric substrates. A N89D/E92K mutant is more stable but considerably less active than wild type. An N89D mutant is more stable and of similar activity to wild type. The substantial change in activity may be due to direct interaction of residue 92 with aggregated substrate or may be via second calcium binding. Second calcium binding may be more probable as activity against monomers is also affected. Additional calcium binding may therefore be an important way of manipulating the activity of phospholipase A2. Study holds ProTherm entries: 9825, 9826, 9827 Extra Details: electrostatics; alpha-helices; pla2

Submission Details

ID: Y3sZXSFg3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:38 p.m.

Version: 1

Publication Details
Goodenough PW;Bhat KM;Collins ME;Perry BN;Pickersgill RW;Sumner IG;Warwicker J;de Haas GH;Verheij HM,Protein Eng. (1991) Changes in activity of porcine phospholipase A2 brought about by charge engineering of a major structural element to alter stability. PMID:1817255
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1FX9 2000-09-25T00:00:00+0000 2.0 CARBOXYLIC ESTER HYDROLASE COMPLEX (DIMERIC PLA2 + MJ33 INHIBITOR + SULPHATE IONS)
1FXF 2000-09-25T00:00:00+0000 1.85 CARBOXYLIC ESTER HYDROLASE COMPLEX (DIMERIC PLA2 + MJ33 INHIBITOR + PHOSPHATE IONS)
1HN4 2000-12-06T00:00:00+0000 1.5 PROPHOSPHOLIPASE A2 DIMER COMPLEXED WITH MJ33, SULFATE, AND CALCIUM
1L8S 2002-03-21T00:00:00+0000 1.55 CARBOXYLIC ESTER HYDROLASE COMPLEX (DIMERIC PLA2 + LPC-ether + ACETATE + PHOSPHATE IONS)
1P2P 1983-06-27T00:00:00+0000 2.6 STRUCTURE OF PORCINE PANCREATIC PHOSPHOLIPASE A2 AT 2.6 ANGSTROMS RESOLUTION AND COMPARISON WITH BOVINE PHOSPHOLIPASE A2
1PIR 1994-12-22T00:00:00+0000 0 SOLUTION STRUCTURE OF PORCINE PANCREATIC PHOSPHOLIPASE A2
1PIS 1994-12-22T00:00:00+0000 0 SOLUTION STRUCTURE OF PORCINE PANCREATIC PHOSPHOLIPASE A2
1SFV 1996-02-20T00:00:00+0000 0 PORCINE PANCREAS PHOSPHOLIPASE A2, NMR, MINIMIZED AVERAGE STRUCTURE
1SFW 1996-02-23T00:00:00+0000 0 PORCINE PANCREAS PHOSPHOLIPASE A2, NMR, 18 STRUCTURES
1Y6O 2004-12-06T00:00:00+0000 2.0 Crystal structure of disulfide engineered porcine pancreatic phospholipase A2 to group-X isozyme in complex with inhibitor MJ33 and phosphate ions

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
96.8 Phospholipase A2, major isoenzyme P04416 PA22_PIG
100.0 Phospholipase A2, major isoenzyme P00592 PA21B_PIG