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
1SFW 1996-07-11 PORCINE PANCREAS PHOSPHOLIPASE A2, NMR, 18 STRUCTURES
1PIS 1995-06-03 SOLUTION STRUCTURE OF PORCINE PANCREATIC PHOSPHOLIPASE A2
1PIR 1995-06-03 SOLUTION STRUCTURE OF PORCINE PANCREATIC PHOSPHOLIPASE A2
1SFV 1996-07-11 PORCINE PANCREAS PHOSPHOLIPASE A2, NMR, MINIMIZED AVERAGE STRUCTURE
1HN4 2001-12-05 1.5 PROPHOSPHOLIPASE A2 DIMER COMPLEXED WITH MJ33, SULFATE, AND CALCIUM
1L8S 2002-12-25 1.55 CARBOXYLIC ESTER HYDROLASE COMPLEX (DIMERIC PLA2 + LPC-ether + ACETATE + PHOSPHATE IONS)
1FXF 2001-09-25 1.85 CARBOXYLIC ESTER HYDROLASE COMPLEX (DIMERIC PLA2 + MJ33 INHIBITOR + PHOSPHATE IONS)
2B00 2006-11-14 1.85 Crystal Structure of Porcine Pancreatic Phospholipase A2 in Complex with Glycocholate
2AZY 2006-11-14 1.9 Crystal Structure of Porcine Pancreatic Phospholipase A2 in Complex with Cholate
1FX9 2001-09-25 2.0 CARBOXYLIC ESTER HYDROLASE COMPLEX (DIMERIC PLA2 + MJ33 INHIBITOR + SULPHATE IONS)
1Y6O 2005-03-22 2.0 Crystal structure of disulfide engineered porcine pancreatic phospholipase A2 to group-X isozyme in complex with inhibitor MJ33 and phosphate ions
3P2P 1990-01-15 2.1 ENHANCED ACTIVITY AND ALTERED SPECIFICITY OF PHOSPHOLIPASE A2 BY DELETION OF A SURFACE LOOP
2AZZ 2006-11-14 2.2 Crystal Structure of Porcine Pancreatic Phospholipase A2 in Complex with Taurocholate
2B01 2006-11-14 2.2 Crystal Structure of Porcine Pancreatic Phospholipase A2 in Complex with Taurochenodeoxycholate
2PHI 1993-07-15 2.2 A LARGE CONFORMATIONAL CHANGE IS FOUND IN THE CRYSTAL STRUCTURE OF THE PORCINE PANCREATIC PHOSPHOLIPASE A2 POINT MUTANT F63V
1Y6P 2005-03-22 2.25 Crystal structure of disulfide engineered porcine pancratic phospholipase a2 to group-x isozyme
2B03 2006-11-14 2.3 Crystal Structure of Porcine Pancreatic Phospholipase A2 in Complex with Taurochenodeoxycholate
4DBK 2012-01-25 2.3 Crystal structure of porcine pancreatic phospholipase A2 complexed with berberine
3FVJ 2010-03-16 2.3 Crystal structure of phospholipase A2 1B crystallized in the presence of octyl sulfate
4G5I 2012-11-07 2.4 Crystal Structure of Porcine pancreatic PlA2 in complex with DBP
4P2P 1992-01-15 2.4 AN INDEPENDENT CRYSTALLOGRAPHIC REFINEMENT OF PORCINE PHOSPHOLIPASE A2 AT 2.4 ANGSTROMS RESOLUTION
3L30 2010-01-26 2.4 Crystal structure of porcine pancreatic phospholipase A2 complexed with dihydroxyberberine
5P2P 1991-10-15 2.4 X-RAY STRUCTURE OF PHOSPHOLIPASE A2 COMPLEXED WITH A SUBSTRATE-DERIVED INHIBITOR
3HSW 2009-06-30 2.5 Crystal Structure of Porcine Pancreatic Phospholipase A2 in Complex with 2-methoxycyclohexa-2-5-diene-1,4-dione
2B04 2006-11-14 2.5 Crystal Structure of Porcine Pancreatic Phospholipase A2 in Complex with Glycochenodeoxycholate
4O1Y 2014-01-29 2.5 Crystal structure of Porcine Pancreatic Phospholipase A2 in complex with 1-Naphthaleneacetic acid
3QLM 2011-04-06 2.5 Crystal structure of porcine pancreatic phospholipase A2 in complex with n-hexadecanoic acid
3O4M 2010-08-25 2.5 Crystal structure of porcine pancreatic phospholipase A2 in complex with 1,2-dihydroxybenzene
1P2P 1983-09-15 2.6 STRUCTURE OF PORCINE PANCREATIC PHOSPHOLIPASE A2 AT 2.6 ANGSTROMS RESOLUTION AND COMPARISON WITH BOVINE PHOSPHOLIPASE A2
3FVI 2010-03-16 2.7 Crystal Structure of Complex of Phospholipase A2 with Octyl Sulfates

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