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Journal of Lipid Research, Vol 38, 745-758, Copyright © 1997 by Lipid Research, Inc.
JJ Luiken, FA van Nieuwenhoven, G America, GJ van der Vusse and JF Glatz
The precise mechanism of uptake of long-chain fatty acids (FA) by cardiac
myocytes is incompletely understood. We examined the involvement of
sarcolemmal proteins in the initial uptake of FA by isolated rat cardiac
myocytes, and the relation between initial uptake and metabolism. Cardiac
myocytes were incubated in the presence of 90 microns [1-14C]palmitate
complexed to 300 microns bovine serum albumin (BSA), presenting a
physiologically relevant condition. During initial palmitate uptake (3
min), 56% of the intracellularly sequestered palmitate was esterified, and
an additional 21% converted into oxidation intermediates. Varying the
palmitate/BSA molar ratio revealed saturation kinetics with the apparent Km
for cellular palmitate uptake (435 micro M) to be comparable to those for
esterification (465 micro M) and oxidation (222 micro M). Varying the BSA
concentration at a fixed palmitate/BSA molar ratio also showed saturation
of uptake at increasing concentrations, with an apparent Km for BSA of 23
micro M. Changes in palmitate metabolism induced by changes in glucose
utilization were accompanied by identical effects on palmitate uptake.
Addition of lactate also inhibited both oxidation and uptake of palmitate,
but had no effect on esterification. Virtually complete inhibition of
palmitate oxidation by etomoxir inhibited palmitate uptake for 50%, while
decreasing esterification by 33%. In the presence of phloretin and trypsin,
palmitate uptake and metabolism were inhibited 76-88%, and in the presence
of sulfo-N-succinimidyloleate by 53%. It is concluded that a) the bulk of
sarcolemmal palmitate translocation occurs by membrane-associated
FA-binding proteins, most likely assisted by albumin binding proteins
without regulatory function, and b) palmitate uptake is most likely driven
by its rapid intracellular metabolic conversion.
ARTICLES
Uptake and metabolism of palmitate by isolated cardiac myocytes from adult rats: involvement of sarcolemmal proteins
Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands.
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