title: Glyoxylate lowers metabolic ATP in human platelets without altering adenylate energy charge or aggregation.
abstract: Human blood platelets adhere to exposed collagen at the site of vascular injury, initiating a signaling cascade leading to fibrinogen activation, secretion of granules and aggregation, thus producing a stable thrombus. All these steps require metabolic ATP. In this study we have labeled the metabolic pool of ATP with nucleotides, treated platelets with various inhibitors and have monitored their ability to be activated. Incubating platelets with glyoxylate dramatically reduced the ATP level without a change in the adenylate energy charge (AEC). This reduction of ATP did not affect ADP-induced primary or secondary aggregation, whereas glyoxal, methyl glyoxal, or the combination of antimycin plus deoxyglucose reduced both ATP and AEC and inhibited aggregation. The reduction of ATP by glyoxylate was almost quantitatively matched by an increase in hypoxanthine without elevation of ADP. AMP, IMP or inosine, acetoacetate, aspartate, or glutamate had no effect on glyoxylate-induced breakdown of ATP, while pyruvate stopped the ATP reduction fast and efficiently. Glyoxylate also lowered the citrate content. The glyoxylate-induced breakdown of ATP coincided with an increase in fructose-1,6-bisphosphate, indicating that the phosphofructokinase reaction was the main ATP-consuming step. Glyoxylate was a substrate for lactate dehydrogenase although with a Km almost 100 times higher than pyruvate. We suggest that glyoxylate primarily competes with pyruvate in the pyruvate dehydrogenase reaction, thus lowering the citrate concentration, which in turn activates phosphofructokinase. Clearly, lowering of ATP in the cytosol by more than 50% does not affect platelet aggregation provided that the AEC is not reduced.
authors.affiliation: The Department of Biomedicine, University of Bergen , Bergen , Norway.
authors.forename: Carol A
authors.initials: CA
authors.lastname: Dangelmaier
authors.forename: Holm
authors.initials: H
authors.lastname: Holmsen
mesh_terms: D000225:Adenine / Q000097:blood; D000244:Adenosine Diphosphate / Q000097:blood / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000097:blood*; D000262:Adenylyl Cyclases / Q000097:blood; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D002250:Carbon Radioisotopes / Q000097:blood; D004734:Energy Metabolism / Q000187:drug effects; D005260:Female; D006038:Glyoxylates / Q000097:blood* / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D019344:Lactic Acid / Q000097:blood; D008297:Male; D010974:Platelet Aggregation / Q000187:drug effects*; D013917:Thrombin
chemical_list: D002250:Carbon Radioisotopes; D006038:Glyoxylates; D019344:Lactic Acid; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D013917:Thrombin; D000262:Adenylyl Cyclases; D000225:Adenine; C031150:glyoxylic acid