TY - JOUR
T1 - Preparation and Characterization of Human Recombinant Protein 1/Clara Cell Mr 10000 Protein
AU - Okutani, Ryuta
AU - Itoh, Yoshihisa
AU - Yamada, Toshiynki
AU - Yamaguchi, Tetsuji
AU - Kawai, Tadashi
AU - Singh, Gurmukh
AU - Yagisawa, Hitoshi
PY - 1996
Y1 - 1996
N2 - 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.
AB - 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.
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U2 - 10.1515/cclm.1996.34.9.691
DO - 10.1515/cclm.1996.34.9.691
M3 - Article
C2 - 8891520
AN - SCOPUS:0029758197
SN - 1434-6621
VL - 34
SP - 691
EP - 696
JO - Clinical Chemistry and Laboratory Medicine
JF - Clinical Chemistry and Laboratory Medicine
IS - 9
ER -