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Bovine Brain Diacylglycerol Lipase: Substrate Specificity and Activation by Cyclic AMP-dependent Protein Kinase

  • Original Article
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Lipids

Abstract

Diacylglycerol lipase (EC 3.1.1.3) was purified from bovine brain microsomes using multiple column chromatographic techniques. The purified enzyme migrates as a single band on SDS-PAGE and has an apparent molecular weight of 27 kDa. Substrate specificity experiments using mixed molecular species of 1,2-diacyl-sn-glycerols indicate that low concentrations of Ca2+ and Mg2+ have no direct effect on enzymic activity and 1,2-diacyl-sn-glycerols are the preferred substrate over 1,3-diacyl-sn-glycerols. The enzyme hydrolyzes stearate in preference to palmitate from the sn-1 position of 1,2-diacyl-sn-glycerols. 1-O-Alkyl-2-acyl-sn-glycerols are not a substrate for the purified enzyme. The native enzyme had a V max value of 616 nmol/min mg protein. Phosphorylation by cAMP-dependent protein kinase resulted in a threefold increase in catalytic throughput (V max = 1,900 nmol/min mg protein). The substrate specificity and catalytic properties of the bovine brain diacylglycerol lipase suggest that diacylglycerol lipase may regulate protein kinase C activity and 2-arachidonoyl-sn-glycerol levels by rapidly altering the intracellular concentration of diacylglycerols.

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Abbreviations

DAG:

Diacylglycerols

PKC:

Protein kinase C

PKA:

cAMP-dependent protein kinase

PLC:

Phospholipase C

PLD:

Phospholipase D

CB-1:

Cannabinoid receptor 1

2-AG:

2-Arachidonoyl-sn-glycerol

PtdIns(4,5)P2 :

Phosphatidylinositol-4,5-bisphosphate

Ins(1,4,5)P3 :

Inositol-1,4,5-trisphosphate

thioester substrate:

rac-1,2-S,O-didecanoyl-1-mercapto-2,3-propanediol

physiologic substrate:

1-Stearoyl-2-arachidonoyl-sn-glycerol

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Acknowledgments

This work was supported in part by research grants NS-10165 and NS-29441 from the National Institutes of Health, US Public Health Service.

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Authors and Affiliations

  1. Department of Pharmacology, Physiology, and Therapeutics, University of North Dakota, School of Medicine and Health Sciences, 501 North Columbia Road, Rm. 3742A, Grand Forks, ND, 58203, USA

    Thad A. Rosenberger

  2. Department of Molecular and Cellular Biochemistry, The Ohio State University, 1645 Neil Avenue, Columbus, OH, 43210, USA

    Akhlaq A. Farooqui & Lloyd A. Horrocks

Authors
  1. Thad A. Rosenberger
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  2. Akhlaq A. Farooqui
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  3. Lloyd A. Horrocks
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Correspondence to Thad A. Rosenberger.

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Rosenberger, T.A., Farooqui, A.A. & Horrocks, L.A. Bovine Brain Diacylglycerol Lipase: Substrate Specificity and Activation by Cyclic AMP-dependent Protein Kinase . Lipids 42, 187–195 (2007). https://doi.org/10.1007/s11745-007-3019-7

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  • DOI: https://doi.org/10.1007/s11745-007-3019-7

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