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Purification and characterization of phospholipase A2 isoforms from the hepatopancreas of red sea bream, Pagrus major1.
- Source :
- Fish Physiology & Biochemistry; Mar1998, Vol. 18 Issue 2, p135-147, 13p
- Publication Year :
- 1998
-
Abstract
- Two phospholipase A<subscript>2</subscript> (PLA<subscript>2</subscript>) isoforms, tentatively denoted as DE-1 and DE-2 PLA<subscript>2</subscript>s, were purified from the hepatopancreas of red sea bream (Pagrus major) to near homogeneity by sequential column chromatography on S-Sepharose fast flow, DEAE-Sepharose fast flow and butyl-Cellulofine, and by ion-exchange, gel-filtration and reversed-phase HPLC. The purified DE-1 and DE-2 PLA<subscript>2</subscript>s both showed a single band with the apparent molecular mass of approx. 13.5 kDa by SDS-polyacrylamide gel electrophoresis, and were found to be both related to group I PLA<subscript>2</subscript> based on the N-terminal amino acid sequences. DE-1 PLA<subscript>2</subscript> had a pH optimum in the alkaline region at around pH 10 and required approximately 10 mM of Ca<superscript>2+</superscript> and 4-10 mM of sodium deoxycholate for its maximal activity, using 2 mM of phosphatidylcholine and phosphatidylethanolamine as substrates. DE-2 PLA<subscript>2</subscript> also had a pH optimum in the alkaline region at around pH 8-9 and required >10 mM of Ca<superscript>2+</superscript> and approximately 6 mM of sodium deoxycholate for its maximum activity with 2 mM of phosphatidylcholine as a substrate; its enzymatic activity towards phosphatidylethanolamine was greatly inhibited by the addition of sodium deoxycholate. The results demonstrate that red sea bream hepatopancreas contains two enzymatically distinct group I PLA<subscript>2</subscript> isoforms. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 09201742
- Volume :
- 18
- Issue :
- 2
- Database :
- Complementary Index
- Journal :
- Fish Physiology & Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 50028842
- Full Text :
- https://doi.org/10.1023/A:1007750618685