Macrobrachium borellii hepatopancreas contains a mitochondrial glycerol-3-phosphate acyltransferase which initiates triacylglycerol biosynthesis.
Mammals express four isoforms of glycerol-3-phosphate acyltransferase (GPAT). The mitochondrial isoform GPAT1 may have been the acyltransferase that appeared first in evolution. The hepatopancreas of the crustacean Macrobrachium borellii has a high capacity for triacylglycerol (TAG) biosynthesis and storage. In order to understand the mechanism of glycerolipid biosynthesis in M. borellii, we investigated its hepatopancreas GPAT activity. In hepatopancreas mitochondria, we identified a GPAT activity with characteristics similar to those of mammalian GPAT1. The activity was resistant to inactivation by SH-reactive N-ethylmaleimide, it was activated by polymyxin-B, and its preferred substrate was palmitoyl-CoA. The reaction products were similar to those of mammalian GPAT1. A 70-kDa protein band immunoreacted with an anti-rat liver GPAT1 antibody. Surprisingly, we did not detect high GPAT specific activity in hepatopancreas microsomes. GPAT activity in microsomes was consistent with mitochondrial contamination, and its properties were similar to those of the mitochondrial activity. In microsomes, TAG synthesis was not dependent on the presence of glycerol-3 phosphate as a substrate, and the addition of monoacylglycerol as a substrate increased TAG synthesis 2-fold. We conclude that in M. borellii the de novo triacylglycerol biosynthetic pathway can be completed in the mitochondria. In contrast, TAG synthesis in the ER may function via the monoacylglycerol pathway.
Pellon-Maison
M
M
Instituto de Investigaciones Bioquímicas de La Plata, CCT La Plata, CONICET INIBIOLP, Facultad Ciencias Médicas UNLP, Calles 60 & 120, La Plata, Argentina.
Garcia
C F
CF
Cattaneo
E R
ER
Coleman
R A
RA
Gonzalez-Baro
M R
MR
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004951:Esterification; D005992:Glycerol-3-Phosphate O-Acyltransferase / Q000378:metabolism*; D043143:Hepatopancreas / Q000201:enzymology*; D008862:Microsomes, Liver / Q000378:metabolism; D008930:Mitochondria, Liver / Q000201:enzymology*; D033541:Palaemonidae; D014280:Triglycerides / Q000096:biosynthesis*
D014280:Triglycerides; D005992:Glycerol-3-Phosphate O-Acyltransferase