Uptake and metabolism of palmitate by isolated cardiac myocytes from adult rats: involvement of sarcolemmal proteins.
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.
Luiken
J J
JJ
Department of Physiology, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, The Netherlands.
van Nieuwenhoven
F A
FA
America
G
G
van der Vusse
G J
GJ
Glatz
J F
JF
D000818:Animals; D001692:Biological Transport; D002417:Cattle; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D008297:Male; D009124:Muscle Proteins / Q000502:physiology*; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D010168:Palmitates / Q000378:metabolism* / Q000494:pharmacology; D051381:Rats; D011917:Rats, Inbred Lew; D012508:Sarcolemma / Q000378:metabolism*; D012709:Serum Albumin / Q000494:pharmacology; D013997:Time Factors
D009124:Muscle Proteins; D010168:Palmitates; D012709:Serum Albumin