Purification and immunological determination of α2-macroglobulin in serum from injured rats

H. Okubo, O. Miyanaga, M. Nagano, H. Ishibashi, J. Kudo, T. Ikuta, K. Shibata

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59 Scopus citations

Abstract

Rat α2-macroglobulin was isolated and purified from the pooled sera of turpentine-injected rats by sequential use of dextran sulphate, DEAE-cellulose, and gel filtration chromatography. The final protein product obtained by this procedure proved to be α2-macroglobulin of a high degree of purity based on electrophoretic, immunologic and centrifugal analysis. The α2-macroglobulin preparation also binds stoichiometrically to trypsin preventing subsequent inhibition by protein trypsin inhibitors. SDS-polyacrylamide gel electrophoresis of rat α2-macroglobulin after incubation with trypsin suggested that there are at least two susceptible peptide bonds in the 170 000-dalton α2-macroglobulin subunit. The concentration of α2-macroglobulin in the sera of rats was measured by electroimmuno assay using a monospecific antiserum against α2-macroglobulin. Purified α2-macroglobulin was used as a standard. Sera from normal male rats contained 32 ± 4 μg of α2-macroglobulin per ml. To determine the time course of response of α2-macroglobulin to inflammation, rats were subjected to either laparotomy or subcutaneous injection of turpentine. After either type of injury, the concentration of α2-macroglobulin increased rapidly, reaching a maximum value of 110-140 times that of the control value by 24 h. Little difference was noted in responsiveness between the two sexes.

Original languageEnglish (US)
Pages (from-to)257-267
Number of pages11
JournalBBA - Protein Structure
Volume668
Issue number2
DOIs
StatePublished - Apr 28 1981

Keywords

  • Acute phase protein
  • Immunological determination
  • Inflammation
  • Protease inhibitor
  • α-Macroglobulin

ASJC Scopus subject areas

  • Medicine(all)

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