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Recombinant human prorenin (rh-prorenin) was purified from supernatants of Chinese hamster ovary (CHO) cell line transfected with the cDNA for rh-prorenin by employing a simple two-step procedure which consisted of ammonium sulfate precipitation and immunoaffinity chromatography using a monoclonal antibody specific for the profragment of human prorenin. About 100-fold purification with 35% recovery was achieved after the two steps. Purified rh-prorenin migrated as a single protein band with apparent molecular weights of 46,000-47,000 and about 50,000 on SDS-PAGE and gel filtration (HPLC), respectively, although it consisted of multiple components (pI values, 5.6-6.4) that could be resolved by isoelectric focusing (IEF). The treatment of rh-prorenin with endo-beta-N-acetylglucosaminidase converted the rather broad protein band to a sharp band on SDS-PAGE and reduced the number of multiple pI peaks on IEF. Amino-terminal sequence analysis of both the purified rh-prorenin and rh-renin revealed Leu-Pro-Thr-Asp- and Leu-Thr-Leu-Gly-, respectively, which agreed with those predicted from the base sequences of their cDNA. These data suggested that microheterogeneity of rh-prorenin is due to the carbohydrate moiety, but not to the protein moiety. Purified rh-prorenin was almost inactive, but was cleaved at the carboxyl end of a dibasic pair Lys-2-Arg-1 by trypsin and converted to active renin. However, at the early stage during trypsin activation, new intermediate forms between rh-prorenin and rh-renin were formed, suggesting multiple activation steps of rh-prorenin in addition to the one step activation.
Y Ishizuka, and
A Shoda, and
S Yoshida, and
Y Kawamura, and
K Haraguchi, and
K Murakami
August 1992,
Biochemistry,
Y Ishizuka, and
A Shoda, and
S Yoshida, and
Y Kawamura, and
K Haraguchi, and
K Murakami
April 1993,
The Journal of biological chemistry,
Y Ishizuka, and
A Shoda, and
S Yoshida, and
Y Kawamura, and
K Haraguchi, and
K Murakami
April 1988,
Biochemical and biophysical research communications,
Y Ishizuka, and
A Shoda, and
S Yoshida, and
Y Kawamura, and
K Haraguchi, and
K Murakami
May 1987,
Biochemistry,
Y Ishizuka, and
A Shoda, and
S Yoshida, and
Y Kawamura, and
K Haraguchi, and
K Murakami
April 1993,
Protein expression and purification,
Y Ishizuka, and
A Shoda, and
S Yoshida, and
Y Kawamura, and
K Haraguchi, and
K Murakami
December 1989,
The Journal of biological chemistry,
Y Ishizuka, and
A Shoda, and
S Yoshida, and
Y Kawamura, and
K Haraguchi, and
K Murakami
March 2000,
Protein expression and purification,
Y Ishizuka, and
A Shoda, and
S Yoshida, and
Y Kawamura, and
K Haraguchi, and
K Murakami
March 1987,
Cancer research,
Y Ishizuka, and
A Shoda, and
S Yoshida, and
Y Kawamura, and
K Haraguchi, and
K Murakami
October 1986,
Proteins,
Y Ishizuka, and
A Shoda, and
S Yoshida, and
Y Kawamura, and
K Haraguchi, and
K Murakami
July 1989,
Journal of biochemistry,
