Homocysteine thiolactone disposal by human arterial endothelial cells and serum in vitro.
Previous work with cultured mammalian cells and perfused laboratory animals suggested to us that hydrolysis of homocysteine thiolactone was catalyzed in these systems. We confirmed this finding by measuring the sulfhydryl-releasing activity of cultured endothelial cells from human umbilical arteries in homocysteine thiolactone solution, pH 7.4, 37 degrees C. The reaction was vigorous and stereospecific and showed saturation kinetics (Km values for L- and D,L-homocysteine thiolactone were 3.9 and 8.2 mmol/l, respectively, and Vmax values were 10.75 and 10.1 mumol/min/10(9) cells, respectively). L-Homocysteine thiolactone was quantitatively converted to homocysteine, as measured by amino acid analysis. Human serum also accelerated the elimination of homocysteine thiolactone, although in this process, the majority of the newly formed sulfhydryl-containing product was precipitable by sulfosalicylic acid, indicating likely homocysteinylation of serum proteins. However, approximately 38% of the sulfhydryl-containing product was not precipitated, and because thiolactone elimination stereospecifically favored the L-enantiomer, a possible subsidiary role for serum-catalyzed hydrolysis of the thiolactone was suggested. No homocysteine thiolactone could be found in serum samples from six patients with acute myocardial infarction, three patients with cystathionine beta-synthase deficiency, and six normal subjects. Thus, humans have active vascular systems for elimination of homocysteine thiolactone, a process that could be responsible for an absence of the compound in serum.
Dudman
N P
NP
Department of Medicine, University of New South Wales, Prince Henry Hospital, Little Bay, Sydney, Australia.
Hicks
C
C
Lynch
J F
JF
Wilcken
D E
DE
Wang
J
J
D001557:Benzenesulfonates; D001769:Blood; D002478:Cells, Cultured; D011232:Chemical Precipitation; D003541:Cystathionine beta-Synthase / Q000172:deficiency; D004730:Endothelium, Vascular / Q000378:metabolism*; D006710:Homocysteine / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D009203:Myocardial Infarction / Q000097:blood; D012459:Salicylates; D013237:Stereoisomerism; D014469:Umbilical Arteries
D001557:Benzenesulfonates; D012459:Salicylates; D006710:Homocysteine; C007957:homocysteine thiolactone; D003541:Cystathionine beta-Synthase; C003366:sulfosalicylic acid