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KMID : 0578319930030010071
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Molecules and Cells
1993 Volume.3 No. 1 p.71 ~ p.74
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Molecular Properties of Recombinant Human α1-Antitrypsin Produced in Escherichia coli and in Vitro Translation System
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Lee, Kee Nyung
Shin, Hwa Soo/Kwon, Ki-sun/Park, Sang Dai/yu, Myeong-hee
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Abstract
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The cDNA encoding human al-antitrypsin (aIAT) was expressed in Escherichia coli upto ---40% of total cellular proteins. The recombinant aIAT carrying the Z type mutation (G1u341--->Lys) were accumulated as aggregates inside the E. coli cells while the wild type alAT were produced as a soluble form. The aggregation caused by the Z mutation became more significant as the growth temperature increased from 30 c to 37 *C. A protein folding assay of alAT on SDS-polyacrylamide gel electrophoresis revealed that the Z polypeptides translated in vitro were in a conformation which could not bind elastase but rather served as a substrate for the protease. The results are consistent with the previously known Z type defects, aggregation inside the hepatocytes of patients and temperature-dependent oligomerization of the purified Z protein. The E. coli expression system and the folding assay of in vitro translation product developed in the present study may be further explored for characterization of other genetic variants of aIAT.
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