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

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted 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 Tyagi, S. R. Articles by Lambeth, J. D. Search for Related Content PubMed PubMed Citation Articles by Tyagi, S. R. Articles by Lambeth, J. D. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 264, Issue 22, 12977-12982, Aug, 1989

On the biological occurrence and regulation of 1-acyl and 1-O-alkyl- diradylglycerols in human neutrophils. Selective destruction of diacyl species using Rhizopus lipase

SR Tyagi, DN Burnham and JD Lambeth
Department of Biochemistry, Emory University Medical School, Atlanta, Georgia 30322.

The occurrence and regulation of 1-ether-linked diradylglycerol in human neutrophils were investigated using a sensitive and practical analytical mass method which distinguishes 1-O-alkyl- (EAG) versus 1- acyl (DAG) diglycerides. After phosphorylation of diglycerides to the corresponding [32P]phosphatidic acids using [gamma-32P]ATP and diglyceride kinase (Preiss, J., Loomis, C. R., Bishop, W. R., Stein, R., Niedel, J. E., and Bell, R. M. (1986) J. Biol. Chem. 261, 8597- 8600), lipase from Rhizopus arrhizus selectively degraded the 1-acyl- containing species (DAG), but the ether lipid (EAG) was resistant and was identified and quantified after thin layer chromatography separation. By using this method, unstimulated neutrophils were demonstrated to contain both DAG and EAG (100-180 and 40-95 pmol/10(7) cells, respectively). The chemoattractant formyl-methionyl-leucyl- phenylalanine (fMLP) caused a rapid (30 s) and transient increase (1.6- fold) in DAG, but no increase in EAG. Opsonized zymosan produced a 6-8- fold sustained increase in DAG peaking at 2 to 3 min, but only a small (1.7-fold) increase in EAG which was not seen until later times (10 min). Thus, under these stimulation conditions, the major diglyceride was DAG. However, in neutrophils "primed" with cytochalasin B or phorbol ester, formyl-methionyl-leucyl-phenylalanine caused a significant increase in EAG. Neutrophils pretreated with cytochalasin B and then stimulated by fMLP showed a rapid (15-60 s) increase (more than 3-fold) in total diglycerides which was sustained beyond 5 min. At the earliest time points (15-30 s), the increase was due almost entirely to DAG (3-fold), but at 1 min and beyond, EAG comprised as much as 40% of the total (up to a 5-fold increase in EAG). Neutrophils pretreated with phorbol ester prior to fMLP stimulation showed a rapid (around 30 s) more than 2-fold increase in both DAG and EAG. Thus, priming conditions (in particular cytochalasin B) may alter either the access of phospholipase(s) C and/or D to membrane phospholipids or may affect their activities, allowing hydrolysis of 1-O-alkyl-containing lipids to generate 1-O-alkyl-containing diglycerides.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. Karima, A. Kantarci, T. Ohira, H. Hasturk, V. L. Jones, B-H. Nam, A. Malabanan, P. C. Trackman, J. A. Badwey, and T. E. Van Dyke Enhanced superoxide release and elevated protein kinase C activity in neutrophils from diabetic patients: association with periodontitis J. Leukoc. Biol., October 1, 2005; 78(4): 862 - 870. [Abstract] [Full Text] [PDF]

				E. M.B. Raeder, P. J. Mansfield, V. Hinkovska-Galcheva, L. Kjeldsen, J. A. Shayman, and L. A. Boxer Sphingosine Blocks Human Polymorphonuclear Leukocyte Phagocytosis Through Inhibition of Mitogen-Activated Protein Kinase Activation Blood, January 15, 1999; 93(2): 686 - 693. [Abstract] [Full Text] [PDF]

				M. E. Venable, A. Bielawska, and L. M. Obeid Ceramide Inhibits Phospholipase D in a Cell-free System J. Biol. Chem., October 4, 1996; 271(40): 24800 - 24805. [Abstract] [Full Text] [PDF]

				V. A. Sciorra and L. W. Daniel Phospholipase D-derived Products in the Regulation of 12-O-Tetradecanoylphorbol-13-acetate-stimulated Prostaglandin Synthesis in Madin-Darby Canine Kidney Cells J. Biol. Chem., June 14, 1996; 271(24): 14226 - 14232. [Abstract] [Full Text] [PDF]