The complete amino acid sequence of an aspartic protease from Rhizopus chinensis, rhizopuspepsin isozyme pI 5, has been determined. Partial sequences were first obtained from the isolated isozyme by a combination of chemical and proteolytic enzyme cleavages, peptide purifications, and Edman degradations. About one-half of the sequence was revealed by this approach. To complete the amino acid sequence, a cDNA library of R. chinensis in pBR322 was constructed. An oligonucleotide probe was synthesized based on the sequence Trp-Trp-Gly-Ile-Thr, and about 40 positive clones were identified by colony hybridization. A clone, 33E2, which had an insert size of about 1.1 kilobase pairs, was found to contain the entire coding region of rhizopuspepsin isozyme pI 5. The sequence of rhizopuspepsin contains 325 amino acid residues. The alignment of the rhizopuspepsin sequence against other aspartic proteases revealed expected homology, with the closest similarity to penicillopepsin which shares 39% identical residues. Porcine pepsin shares about 36% identical residues with rhizopuspepsin.
|Number of pages||7|
|Journal||Journal of Biological Chemistry|
|Publication status||Published - 5 Feb 1987|
Scopus Subject Areas
- Molecular Biology
- Cell Biology