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. 1990 Nov 1;271(3):565-9.
doi: 10.1042/bj2710565.

Rapid and sensitive techniques for identification and analysis of 'reactive-centre' mutants of C1-inhibitor proteins contained in type II hereditary angio-oedema plasmas

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Rapid and sensitive techniques for identification and analysis of 'reactive-centre' mutants of C1-inhibitor proteins contained in type II hereditary angio-oedema plasmas

K S Aulak et al. Biochem J. .

Abstract

Novel procedures for structural analysis of the 'reactive-centre' residues, particularly the P1 residue, of the dysfunctional C1-inhibitor proteins found in the plasmas of type II hereditary angio-oedema (HAE) patients are described. C1-inhibitor is adsorbed directly from plasma on to Sepharose-anti-(C1 inhibitor) beads. The P1 residue of C1 inhibitor is arginine and hence a potential cleavage site for trypsin. Thus trypsin digestion of the immobilized protein, followed by SDS/PAGE of the released fragments, identifies P1 residue mutations. Pseudomonas aeruginosa elastase digestion of the immobilized protein, followed by purification of the released C-terminal peptide (by h.p.l.c.) and N-terminal sequence analysis defines the new P1 residue (or other mutations in the reactive-centre region). The techniques are both rapid and highly sensitive, requiring only 400 microliters of plasma. In addition, they permit accurate assessment of the level of normal (functional) inhibitor in a subclass of type II HAE plasmas, those containing P1-residue mutant proteins.

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References

    1. Biochem Biophys Res Commun. 1967 Apr 20;27(2):157-62 - PubMed
    1. Am J Med. 1963 Jul;35:37-44 - PubMed
    1. Nature. 1970 Aug 15;227(5259):680-5 - PubMed
    1. J Lab Clin Med. 1970 Nov;76(5):809-15 - PubMed
    1. J Clin Invest. 1971 Oct;50(10):2143-9 - PubMed

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