Proteolysis of human ceruloplasmin. Some peptide bonds are particularly susceptible to proteolytic attack

K. A. Moshkov, S. Lakatos, J. Hajdu, P. Závodsky, S. A. Neifakh

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Highly purified human ceruloplasmin was isolated from fresh donor blood in the presence of inhibitors of proteolysis and from stored retroplacental blood serum both with and without inhibitors of proteolysis. According to the data of electrophoresis, ultracentrifuge sedimentation velocity and sedimentation equilibrium, all the ceruplasmin samples were homogeneous, their molecular weight being 130000. The dissociation of the samples treated by dodecylsulphate, guanidine HCl, and urea was studied by means of quantitative analytical and preparative electrophoresis, and sedimentation equilibrium. The dissociation patterns depended on whether inhibitors were used in the isolation procedure. Polypeptides with molecular weights of 130000, 112000 and 16000 (minor component) were obtained, if phenylmethylsulfonyl fluoride and/or 6-aminohexanoic acid were used; if these compounds were not used, the dissociation yielded additional polypeptides of molecular weights of 100000 (minor component), 64000 and 48000. Under proteolysis-favouring conditions the relative amount of these polypeptides increased. Prolonged storage of samples under sterile conditions without inhibitors of proteolysis resulted in a decrease of the relative amount of polypeptides with molecular weights of 130000, 112000, 100000, 64000 and 48000, coronary to that of the 16000-M-polypeptide, which increased. At the same time new polypeptides appeared with molecular weights of 42000 and 21500-23000. Spontaneous specific fragmentation of the ceruloplasmin molecule is due to trace amounts of proteases, which seem to originate from bloodplasma. Limited tryptic hydrolysis of the ceruloplasmin globule resulted in the appearance of polypeptides with the same molecular weights which were observed in spontaneous fragmentation. A conclusion is drawn that the ceruloplasmin molecule in vivo is a single polypeptide chain with at least five bonds which in vitro are the points of specific proteolytic fragmentation, yielded six principal fragments.

Original languageEnglish
Pages (from-to)127-134
Number of pages8
JournalEuropean Journal of Biochemistry
Volume94
Issue number1
DOIs
Publication statusPublished - 1979

Fingerprint

Proteolysis
Ceruloplasmin
Peptides
Molecular Weight
Molecular weight
Sedimentation
Electrophoresis
Blood
Phenylmethylsulfonyl Fluoride
Aminocaproic Acid
Molecules
Guanidine
Blood Donors
Urea
Hydrolysis
Peptide Hydrolases

ASJC Scopus subject areas

  • Biochemistry

Cite this

Proteolysis of human ceruloplasmin. Some peptide bonds are particularly susceptible to proteolytic attack. / Moshkov, K. A.; Lakatos, S.; Hajdu, J.; Závodsky, P.; Neifakh, S. A.

In: European Journal of Biochemistry, Vol. 94, No. 1, 1979, p. 127-134.

Research output: Contribution to journalArticle

Moshkov, K. A. ; Lakatos, S. ; Hajdu, J. ; Závodsky, P. ; Neifakh, S. A. / Proteolysis of human ceruloplasmin. Some peptide bonds are particularly susceptible to proteolytic attack. In: European Journal of Biochemistry. 1979 ; Vol. 94, No. 1. pp. 127-134.
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