Purification and characterization of a catalytic domain of rat intestinal phospholipase B/lipase associated with brush border membranes.

The Journal of Biological Chemistry
H TojoM Okamoto

Abstract

A brush border membrane-associated phospholipase B/lipase was solubilized from the distal two-thirds of rat small intestine by autolysis during storage at -35 degrees C over 1 month, and then the enzyme was purified to homogeneity and characterized enzymatically and structurally. The purified enzyme exhibited broad substrate specificity including esterase, phospholipase A2, lysophospholipase, and lipase activities. SDS-gel electrophoretic and reverse-phase high performance liquid chromatographic analyses demonstrated that a single enzyme catalyzes these activities. It preferred hydrolysis at the sn-2 position of diacylphospholipid and diacylglycerol without strict stereoselectivity, whereas it apparently exhibited no positional specificity toward triacylglycerol. Diisopropyl fluorophosphate, an irreversible inhibitor of serine esterases and lipases inhibited purified enzyme. When the position of enzyme on SDS-gel electrophoresis under the non-reducing conditions was determined by assaying the activity eluted from sliced gels, brush border membrane-associated enzyme corresponded to a approximately 150-kDa protein; autolysis gave a 35-kDa product, in agreement with the results of immunoblot analysis. The purified 35-kDa enzyme co...Continue Reading

References

Nov 12, 1975·Journal of Chromatography·V E VaskovskyI M Vasendin
May 1, 1991·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·S Pind, A Kuksis
Jan 1, 1991·Methods in Enzymology·H TojoM Okamoto
Dec 14, 1990·Science·D L ScottP B Sigler
May 15, 1981·Analytical Biochemistry·G L Jensen, A Bensadoun
Nov 18, 1980·Biochimica Et Biophysica Acta·H HauserD E Bowyer
Jan 1, 1981·Methods in Enzymology·L M Pinkus
Dec 1, 1982·The Biochemical Journal·S M ParkinD S Robinson
Oct 27, 1995·The Journal of Biological Chemistry·L W TjoelkerP W Gray
May 1, 1995·Trends in Biochemical Sciences·C Upton, J T Buckley
Aug 29, 1995·Proceedings of the National Academy of Sciences of the United States of America·J BalsindeE A Dennis
Mar 15, 1994·Biochemistry·K ThirstrupF Carrière
Jan 28, 1993·Nature·M J Berridge
Mar 28, 1997·The Journal of Biological Chemistry·J TangS S Jones
Jul 1, 1964·Archives of Biochemistry and Biophysics·G V VAHOUNYC R TREADWELL
May 1, 1992·Trends in Cardiovascular Medicine·R W Gross

❮ Previous
Next ❯

Citations

Jul 9, 1999·Biochimica Et Biophysica Acta·A Wang, E A Dennis
Jun 24, 2011·Chemical Reviews·Jonathan Z Long, Benjamin F Cravatt
Jun 9, 2011·Journal of Agricultural and Food Chemistry·Manami InoueAkira Tokumura
Mar 11, 2010·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Eric D LabontéDavid Y Hui
Jul 27, 1999·Lipids·B ChaminadeH Chap
Nov 21, 2008·The FEBS Journal·Shengyuan XuPer Venge
Aug 27, 2013·Biochemical and Biophysical Research Communications·Shohei KuwataNobuyuki Fukushima
Jul 14, 2007·Journal of Lipid Research·Akira AbeJames A Shayman
Dec 24, 2018·American Journal of Physiology. Gastrointestinal and Liver Physiology·Åke Nilsson, Rui-Dong Duan
Aug 15, 2013·The Journal of Biological Chemistry·Atsushi AsanoAlexander J Travis
Nov 4, 2006·Biochimica Et Biophysica Acta·Gerwald A KöhlerSantosh Nigam

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.