Ribonuclease inhibitor protein of human erythrocytes: characterization, loss of activity in response to oxidative stress, and association with Heinz bodies.
Significant amounts of ribonuclease inhibitor protein are present in human and rat erythrocytes, cells that are essentially devoid of ribonuclease or functional RNA. The protein from human erythrocytes is indistinguishable from human placental ribonuclease inhibitor protein by immunological and biochemical criteria. Each inhibitor forms an equimolar complex with bovine pancreatic ribonuclease A and is inactivated by treatment with the sulfhydryl reagent p-(hydroxymercuri)benzoate. Amino acid composition and several cycles of amino acid sequence analysis also showed apparent identify of the erythrocyte and placental proteins. We calculate a level of 1.5-3.5 x 10(4) molecules of active inhibitor per erythrocyte, most or all of which occurs in an uncomplexed form since inactivation of the inhibitor revealed barely detectable levels of RNase activity. Immunogold localization showed a high level of labeling and a uniform distribution of gold particles in the cytoplasm of erythrocytes, while little inhibitor activity was found in association with isolated red blood cell membranes. Oxidative stress on isolated red cells resulted in a decrease in the level of reduced glutathione and a gradual and irreversible loss of inhibitor activity; inhibitor disappeared from the cytosol and became associated with nascent Heinz bodies. We suggest a role for this protein in the metabolism and aging process of the erythrocyte.
Moenner
M
M
INSERM Unité 427, Développement Humain, Croissance et Différenciation, Faculté des Sciences Pharmaceutiques et Biologiques de Paris, France.
Vosoghi
M
M
Ryazantsev
S
S
Glitz
D G
DG
D000818:Animals; D002417:Cattle; D004912:Erythrocytes / Q000737:chemistry* / Q000201:enzymology / Q000648:ultrastructure; D005260:Female; D005978:Glutathione / Q000378:metabolism; D006366:Heinz Bodies / Q000737:chemistry; D006801:Humans; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D007150:Immunohistochemistry; D008970:Molecular Weight; D009928:Organ Specificity; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010920:Placenta / Q000737:chemistry; D051381:Rats; D012259:Ribonuclease, Pancreatic / Q000037:antagonists & inhibitors; D013045:Species Specificity
D006902:Hydroxymercuribenzoates; C030601:4-hydroxymercuribenzoate; D012259:Ribonuclease, Pancreatic; D005978:Glutathione