KMID : 0545119990090050679
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Journal of Microbiology and Biotechnology 1999 Volume.9 No. 5 p.679 ~ p.682
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Endoproteolytic Processing of Human Parathyroid Hormone in Saccharomyces cerevisiae Mutants Lacking Genes Encoding the Yap3,Mkc7,and Kex2 Endoproteases
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CHOI, WON-A
KANG, HYUN AH/CHUNG, BONG HYUN
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Abstract
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When the human parathyroid hormone (hPTH) is expressed as a secretory product in S. cerevisiae, most of the secreted hPTH is internally cleaved by endoproteolytic processing. To investigate whether the yeast endoproteases such as Kex2p, Yap3p, and Mkc7p are involved in the endoproteolytic processing of hPTH in S. cerevisiae, hPTH was expressed in S. cerevisiae mutants deficient in one or two of the following well-known endoproteases such as Kex2p, Mkc7p, and Yap3p. Among these mutants, the yap3-disrupted (yap3¥Ä) and yap3/mkc7-disrupted (yap3¥Ämkc7¥Ä) yeasts showed a significant reduction in the extent of hPTH proteolysis. In contrast, the mkc7-disrupted (mkc7¥Ä) yeast did not reduce the proteolysis of hPTH as compared to the wild type. This suggests that Mkc7p is not involved in the endoproteolytic processing of hPTH. It was also found that the kex2-disrupted (kex2¥Ä) mutant was not able to secrete a detectable amount of hPTH.
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