HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Tojo, H. Articles by Okamoto, M. Search for Related Content PubMed PubMed Citation Articles by Tojo, H. Articles by Okamoto, M. Social Bookmarking                      What's this?

Vol. 273, Issue 4, 2214-2221, January 23, 1998

Purification and Characterization of a Catalytic Domain of Rat Intestinal Phospholipase B/Lipase Associated with Brush Border Membranes

From the Department of Molecular Physiological Chemistry, Osaka University Medical School, 2-2 Yamadaoka, Suita, Osaka 565, Japan

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 °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 ~150-kDa protein; autolysis gave a 35-kDa product, in agreement with the results of immunoblot analysis. The purified 35-kDa enzyme consisted of a 14-kDa peptide and a glycosylated 21-kDa peptide. Their NH₂-terminal amino acid sequences were determined and found in the second repeat of 161-kDa phospholipase B/lipase with 4-fold tandem repeats of ~38 kDa each, which we cloned and sequenced in the accompanying paper (Takemori, H., Zolotaryov, F., Ting, L., Urbain, T., Komatsubara, T., Hatano, O., Okamoto, M., and Tojo, H. (1998) J. Biol. Chem. 273, 2222-2231). These results indicate that the purified enzyme is the catalytic domain derived from the second repeat of brush border membrane-associated phospholipase B/lipase.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				A. Abe, M. Hiraoka, and J. A. Shayman The acylation of lipophilic alcohols by lysosomal phospholipase A2 J. Lipid Res., October 1, 2007; 48(10): 2255 - 2263. [Abstract] [Full Text] [PDF]

				E. D. Labonte, R. J. Kirby, N. M. Schildmeyer, A. M. Cannon, K. W. Huggins, and D. Y. Hui Group 1B Phospholipase A2-Mediated Lysophospholipid Absorption Directly Contributes to Postprandial Hyperglycemia. Diabetes, April 1, 2006; 55(4): 935 - 941. [Abstract] [Full Text] [PDF]

				D. Y. Hui and P. N. Howles Carboxyl ester lipase: structure-function relationship and physiological role in lipoprotein metabolism and atherosclerosis J. Lipid Res., December 1, 2002; 43(12): 2017 - 2030. [Abstract] [Full Text] [PDF]

				M. Ito, U. Tchoua, M. Okamoto, and H. Tojo Purification and Properties of a Phospholipase A2/Lipase Preferring Phosphatidic Acid, Bis(monoacylglycerol) Phosphate, and Monoacylglycerol from Rat Testis J. Biol. Chem., November 8, 2002; 277(46): 43674 - 43681. [Abstract] [Full Text] [PDF]

				M. Nauze, L. Gonin, B. Chaminade, C. Peres, F. Hullin-Matsuda, B. Perret, H. Chap, and A. Gassama-Diagne Guinea Pig Phospholipase B, Identification of the Catalytic Serine and the Proregion Involved in Its Processing and Enzymatic Activity J. Biol. Chem., November 8, 2002; 277(46): 44093 - 44099. [Abstract] [Full Text] [PDF]

				K. Bhardwaj, A. Raju, and R. Rajasekharan Identification, Purification, and Characterization of a Thermally Stable Lipase from Rice Bran. A New Member of the (Phospho) Lipase Family Plant Physiology, December 1, 2001; 127(4): 1728 - 1738. [Abstract] [Full Text] [PDF]

				H. Takemori, F. N. Zolotaryov, L. Ting, T. Urbain, T. Komatsubara, O. Hatano, M. Okamoto, and H. Tojo Identification of Functional Domains of Rat Intestinal Phospholipase B/Lipase. ITS cDNA CLONING, EXPRESSION, AND TISSUE DISTRIBUTION J. Biol. Chem., January 23, 1998; 273(4): 2222 - 2231. [Abstract] [Full Text] [PDF]