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

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

Research output: Contribution to journalArticle

59 Citations (Scopus)

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
Externally publishedYes

Fingerprint

Macroglobulins
Purification
Rats
Serum
Turpentine
Trypsin
DEAE-Cellulose
Dextran Sulfate
Trypsin Inhibitors
Subcutaneous Injections
Chromatography
Electrophoresis
Laparotomy
Gel Chromatography
Immune Sera
Polyacrylamide Gel Electrophoresis
Assays
Proteins
Gels

Keywords

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

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Purification and immunological determination of α2-macroglobulin in serum from injured rats. / Okubo, H.; Miyanaga, O.; Nagano, M.; Ishibashi, H.; Kudo, J.; Ikuta, Tohru; Shibata, K.

In: BBA - Protein Structure, Vol. 668, No. 2, 28.04.1981, p. 257-267.

Research output: Contribution to journalArticle

Okubo, H. ; Miyanaga, O. ; Nagano, M. ; Ishibashi, H. ; Kudo, J. ; Ikuta, Tohru ; Shibata, K. / Purification and immunological determination of α2-macroglobulin in serum from injured rats. In: BBA - Protein Structure. 1981 ; Vol. 668, No. 2. pp. 257-267.
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