Abstract
Heterologous radioimmunoassays for a semiquantitative analysis of alpha 1-microglobulin were developed, exploiting the binding between polyclonal rabbit or goat antisera against human, guinea pig, or rat alpha 1-microglobulin and 125I-labeled human, guinea pig, or rat alpha 1-microglobulin. Homologues of this protein were detected in human, guinea pig, Rhesus monkey, rat, mouse, rabbit, goat, horse, and cow serum by inhibition of a set of heterologous radioimmunoassays. Serum proteins were separated by gel chromatography, and fractions were pooled, concentrated, and radiolabeled with 125I. By immunoprecipitation of the radioiodinated serum pools with heterologous anti-alpha 1-microglobulin-sera, and separating the precipitates by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, analogues of alpha 1-microglobulin were isolated from serum of man, guinea pig, Rhesus monkey, rat, mouse, horse, and chicken. The apparent molecular weight of alpha 1-microglobulin was 31,000-32,000 in human and monkey serum and 24,000-26,000 in guinea pig, rat, mouse, horse, and chicken serum. The possibility of an addition of a 5,000-8,000-Da peptide in primate alpha 1-microglobulin is discussed.
Highlights
Tative analysis of al-microglobulin were developed, The apparent molecular weight of human al-microglobulin exploiting the binding between polyclonal rabbit or determined by sodium dodecyl sulfate-polyacrylamide gel goatantiseraagainsthuman,guinea pig, or rat al- electrophoresis is 31,000 [7], which is larger than guinea pig microglobulin and ‘Z61-labeled human, guinea pig, or a,microglobulin from urine [2] and rata,microglobulin from rat a,microglobulin
As a detected in human, guinea pig, Rhesus monkey, rat, first step to study this size difference from an evolutionary opfmrfraooacuttesisieone,ntssrwowafeberbhreieetts,peeropgoooallaoretagd,to,euhdcsoorbrnasycdeeg,inoeaitlmrncadmhtreuodcnom, owaaantsobsdsgeayrryriuasan.pmdhShiieobyrl,iuatabmionendledplsyeozrieundmt tofhfreovimeawpsp,ewavreeernathl admvifeoflederceeumnltaosrnpwsetecraiigetehsd.t the presence and anaof al-microglobulin in with laaI
Human a,microglobulin has been reported to have immunoregulatory functions suppressing the blastogenic effect of some antigens onlymphocytes in vitro and inhibitingthe migration of leukocytes in vitro [5]. a]-microglobulin from allthree species is a glycoprotein, and glycopeptides from human and guinea pig al-microglobulin were shown to possess the inhibitory activity of antigen stimulation of lymphocytes [6]
Summary
Normal rabbit serum was used as a al-Microglobulin is a plasma protein, isolated from the urine of humans (I),guinea pigs [2], and rats [3, 4] with disturbed renal filtration and/or reabsorbtion functions. Immunoprecipitation of a,-Microglobulin from Serum Pools-Monomeric al-microglobulin in serum from man, guinea pig, rat, mouse, Rhesus monkey, horse, and chicken was partly purified by gel chromatography through Sephadex G-200. The serum proteins prepared were radiolabeled with 1251,immunoprecipitated with anti-al-microglobulin sera or, as a control, antiserum against guinea pig albumin, and.
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