Copper inhibits activated protein C: protective effect of human albumin and an analogue of its high-affinity copper-binding site, d-DAHK.
Activated protein C (APC) is useful in the treatment of sepsis. Ischemia and acidosis, which often accompany sepsis, cause the release of copper from loosely bound sites. We investigated (i) whether physiological concentrations of copper inhibit APC anticoagulant activity and (ii) if any copper-induced APC inhibition is reversible by human serum albumin (HSA) or a high-affinity copper-binding analogue of the human albumin N-terminus, d-Asp-d-Ala-d-His-d-Lys (d-DAHK). APC activity after 30 min of incubation with CuCl2 (10 microM) was decreased 26% below baseline. HSA, both alone and when combined with various ratios of CuCl2, increased APC activity significantly above baseline. d-DAHK alone and 2:1 and 4:1 ratios of d-DAHK:CuCl2 also increased APC activity. APC contained 1.4 microM copper, which helps explain the increased APC activity with HSA and d-DAHK alone. These in vitro results indicate that copper inhibits APC activity and that albumin and d-DAHK reverse the copper-induced APC deactivation.
Bar-Or
David
D
Trauma Research Department, Swedish Medical Center, 501 East Hampden Avenue, Englewood, Colorado 80110, USA.
Rael
Leonard T
LT
Winkler
James V
JV
Yukl
Richard L
RL
Thomas
Gregory W
GW
Shimonkevitz
Richard P
RP
D001665:Binding Sites; D003300:Copper / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D009842:Oligopeptides / Q000378:metabolism* / Q000494:pharmacology*; D011485:Protein Binding; D011486:Protein C / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D012709:Serum Albumin / Q000378:metabolism / Q000494:pharmacology*
D004791:Enzyme Inhibitors; D009842:Oligopeptides; D011486:Protein C; D012709:Serum Albumin; C431583:aspartyl-alanyl-histidyl-lysine; D003300:Copper; C029892:cupric chloride