PMID: 9434118Jan 20, 1998Paper

Functional implications of disulfide bond, Cys45-Cys50, in recombinant prochymosin

Biochimica Et Biophysica Acta
Y ZhangK Yang

Abstract

Bovine prochymosin (chymosin) contains three disulfide bonds: Cys45-Cys50, Cys206-Cys210 and Cys250-Cys283 (pepsin numbering). We have demonstrated that Cys250-Cys283 is not intimately involved in the catalytic mechanism of chymosin but is required for the correct refolding of prochymosin. To elucidate the functional implications of another disulfide bond of Cys45-Cys50, these two cysteine residues were replaced separately or simultaneously by site-directed mutagenesis. Like the wild-type prochymosin all the seven mutants generated (C45A, C50A, C45A/C50A, C45D, C50S, C45D/C50S, C45A/C50S) exhibit the activity of autocatalytic activation after refolding, indicating that Cys45-Cys50 is dispensable in prochymosin refolding. Spectroscopic analyses and urea-induced denaturation studies of prochymosin and four mutants tested (C45A, C50A, C45A/C50A, C45D/C50S) show that: (1) they share similar far-UV CD spectra and similar fluorescence emission spectra; (2) mutation results in a perturbance of tryptophan environment and somewhat destabilization of prochymosin conformation. However, quenching studies reveal that the only one tryptophan residue inaccessible to acrylamide is still buried in the mutated molecules. All these results sugges...Continue Reading

References

Mar 1, 1979·European Journal of Biochemistry·V B PedersenB Foltmann
Sep 16, 1992·Biochemical and Biophysical Research Communications·K HuangK Yang
Oct 15, 1990·The Biochemical Journal·R SugrueR B Freedman
May 16, 1988·Biochemical and Biophysical Research Communications·Y TaniyamaM Ikehara
Oct 1, 1971·The Journal of Antibiotics·T AoyagiH Umezawa
Jun 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·J S EmtageP A Lowe
Dec 11, 1981·Science·F Sanger

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Citations

May 7, 1999·The Biochemical Journal·C WeiK Yang
Mar 20, 2012·PloS One·Luís Filipe Costa CastroJonathan Mark Wilson
Apr 20, 2005·FEMS Microbiology Letters·Yurong LiangYuying Zhang

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