Characterization of microsomal diacylglycerol acyltransferase activity from bovine adipose and muscle tissue.
The activity of the triacylglycerol bioassembly enzyme, diacylglycerol acyltransferase (DGAT), was characterized in microsomal fractions prepared from bovine subcutaneous (SC) adipose, intramuscular (IM) adipose, and muscle (pars costalis diaphragmatis) tissue. The activity of DGAT was generally higher from SC adipose tissue than from IM adipose or muscle tissue. The characteristics of DGAT activity from the three bovine tissues resembled the activity characteristics observed in previous studies from various other organisms and tissues; the pH optimum was near neutrality, the activity was almost completely inhibited by pre-incubation with N-ethylmaleimide (NEM), and the enzyme accepted a broad range of acyl-CoAs and sn-1,2-diacylglycerols. In some aspects, the SC adipose tissue DGAT activity was different from the DGAT activity from the other two tissues. The SC adipose tissue DGAT activity was not as susceptible to inhibition by NEM as the enzymes from the two other tissue sources, and it exhibited increased specificity for substrates containing oleoyl moieties. The differences in DGAT properties between the three bovine tissues may account to some extent for the differences in the relative fatty acid composition and the positional distribution of fatty acids in triacylglycerol between bovine tissues. The observed differences in enzymatic properties also support recent biochemical and molecular genetic observations that imply the existence of multiple DGAT genes and/or isoforms.
Lozeman
F J
FJ
Department of Chemistry and Biochemistry, University of Lethbridge, Lethbridge, T1K 3M4, Alberta, Canada.
Middleton
C K
CK
Deng
J
J
Kazala
E C
EC
Verhaege
C
C
Mir
P S
PS
Laroche
A
A
Bailey
D R
DR
Weselake
R J
RJ
D000214:Acyl Coenzyme A / Q000378:metabolism; D000217:Acyltransferases / Q000737:chemistry*; D000273:Adipose Tissue / Q000201:enzymology*; D000818:Animals; D002417:Cattle; D002855:Chromatography, Thin Layer; D051048:Diacylglycerol O-Acyltransferase; D004075:Diglycerides / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005033:Ethylmaleimide / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008861:Microsomes / Q000201:enzymology*; D009132:Muscles / Q000201:enzymology*; D020033:Protein Isoforms; D013379:Substrate Specificity; D013997:Time Factors; D014018:Tissue Distribution
D000214:Acyl Coenzyme A; D004075:Diglycerides; D020033:Protein Isoforms; D000217:Acyltransferases; D051048:Diacylglycerol O-Acyltransferase; D005033:Ethylmaleimide