Preparation and Characterization of Human Recombinant Protein 1/Clara Cell Mr 10000 Protein

Ryuta Okutani, Yoshihisa Itoh, Toshiynki Yamada, Tetsuji Yamaguchi, Tadashi Kawai, Gurmukh Singh, Hitoshi Yagisawa

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Protein 1, which is identical to human Clara cell Mr 104 protein, is a homodimeric, low molecular mass protein (Mr 14000) and an effective inhibitor of phospholipase A2 activity. We have expressed this protein in E. coli and characterized its physiochemical and biological properties. Using a pET expression system, about 1.7 mg of purified recombinant protein 1 was obtained from 250 ml of E. coli culture. The amino-terminal sequence of recombinant protein 1 up to the 20th residue was identical to that of native protein 1 except for an extra methionine at the ammo-terminus. On reversed-phase HPLC, recombinant protein 1 eluted at the same retention time as native protein 1. The dose-response curves of recombinant protein 1 and native protein 1 in an enzyme-linked immunosorbent assay for protein 1 were identical. Recombinant protein 1 inhibited both porcine pancreas and cobra venom phospholipase A2 activities. These results indicated that recombinant protein 1 is structurally and biologically identical to native protein 1. We found that recombinant protein 1 also inhibits phosphatidylinositol-specific phospholipase C activity.

Original languageEnglish (US)
Pages (from-to)691-696
Number of pages6
JournalClinical Chemistry and Laboratory Medicine
Volume34
Issue number9
DOIs
StatePublished - Jan 1 1996
Externally publishedYes

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ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

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