EFFICIENT SECRETION OF HUMAN PARATHYROID-HORMONE BY SACCHAROMYCES-CEREVISIAE

被引：44

作者：

GABRIELSEN, OS ^{[1
]}

REPPE, S ^{[1
]}

SAETHER, O ^{[1
]}

BLINGSMO, OR ^{[1
]}

SLETTEN, K ^{[1
]}

GORDELADZE, JO ^{[1
]}

HOGSET, A ^{[1
]}

GAUTVIK, VT ^{[1
]}

ALESTROM, P ^{[1
]}

OYEN, TB ^{[1
]}

GAUTVIK, KM ^{[1
]}

机构：

[1] UNIV OSLO,INST MED BIOCHEM,N-0316 OSLO 3,NORWAY

来源：

GENE | 1990年 / 90卷 / 02期

关键词：

enzyme assay; KEX2; mating factor α fusion; osteoblast adenylate cyclase; proteolytic processing; Recombinant DNA; yeast;

D O I：

10.1016/0378-1119(90)90188-W

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A cDNA encoding mature human parathyroid hormone (hPTH) was expressed in Saccharomyces cerevisiae, after fusion to the prepro region of yeast mating factor α (MFα). Radioimmunoassay showed high levels of hPTH immunoreactive material in the growth medium (up to 10 μg/ml). More than 95% of the immunoreactive material was found extracellularly as multiple forms of hormone peptides. Three internal cleavage sites were identified in the hPTH molecule. The major cleavage site, after a pair of basic amino acids (aa) (Arg25Lys26↓Lys27), resembles that recognized by the KEX2 gene product on which the MFα expression-secretion system depends. The use of a protease-deficient yeast strain and the addition of high concentrations of aa to the growth medium, however, not only changed the peptide pattern, but also resulted in a significant increase in the yield of intact hPTH (1-84) (more than 20% of the total amount of immunoreactive material). The secreted hPTH (1-84) migrates like a hPTH standard in two different gel-electrophoretic systems, co-elutes with standard hPTH on reverse-phase high-performance liquid chromatography, reacts with two hPTH antibodies raised against different parts of the peptide, has a correct N-terminal aa sequence, and has full biological activity in a hormone-sensitive osteoblast adenylate cyclase assay. © 1990.

引用

页码：255 / 262

页数：8

共 38 条

[1]

ABERSOLD RH, 1986, J BIOL CHEM, V261, P4229

[2]

BEGGS JD, 1981, A BENZON S, V16, P383

[3] SECRETION OF FOREIGN PROTEINS FROM SACCHAROMYCES-CEREVISIAE DIRECTED BY ALPHA-FACTOR GENE FUSIONS [J].