Primary structure of a precursor to the aspartic proteinase from Rhizomucor miehei shows that the enzyme is synthesized as a zymogen - PubMed (original) (raw)
Primary structure of a precursor to the aspartic proteinase from Rhizomucor miehei shows that the enzyme is synthesized as a zymogen
E Boel et al. Proteins. 1986 Dec.
Abstract
In order to characterize the zymogen of the milk-clotting enzyme from Rhizomucor miehei, we constructed a cDNA library on pBR327 in Escherichia coli. Aspartic proteinase-specific recombinants were isolated by colony hybridization to a specific oligonucleotide mixture, and the cDNA sequence corresponding to a precursor form of the enzyme was determined. The deduced amino acid sequence shows that this secreted fungal proteinase is synthesized as a precursor. The first 22 amino acid residues in this precursor constitute a typical signal peptide. The amino acid sequence of the following 47-amino-acid-long prosegment shows homology to the prosegments from both the extracellular and intracellular vertebrate aspartic proteinases, and to the prosegments from the yeast and Mucor pusillus aspartic proteinases as well. These observations suggest that all aspartic proteinases are synthesized with a prosegment and that this prosegment is essential for the correct folding of all the mature enzymes. The active Rhizomucor miehei enzyme consists of 361 amino acid residues with a total molecular weight of 38,701. Clusters of identities around the active site cleft support the assumption that these proteinases have a common folding of their peptide chains. The disulphide bridges were localized in the fungal enzyme, and 2 N-glycosylation sites were identified.
Similar articles
- Crystal structure of the aspartic proteinase from Rhizomucor miehei at 2.15 A resolution.
Yang J, Teplyakov A, Quail JW. Yang J, et al. J Mol Biol. 1997 May 2;268(2):449-59. doi: 10.1006/jmbi.1997.0968. J Mol Biol. 1997. PMID: 9159482 - X-ray analyses of aspartic proteinases. V. Structure and refinement at 2.0 A resolution of the aspartic proteinase from Mucor pusillus.
Newman M, Watson F, Roychowdhury P, Jones H, Badasso M, Cleasby A, Wood SP, Tickle IJ, Blundell TL. Newman M, et al. J Mol Biol. 1993 Mar 5;230(1):260-83. J Mol Biol. 1993. PMID: 8450540 - Cloning and sequence analysis of the glyceraldehyde-3-phosphate dehydrogenase gene from the zygomycetes fungus Rhizomucor miehei.
Vastag M, Kasza Z, Acs K, Papp T, Schwab H, Vágvölgyi C. Vastag M, et al. Antonie Van Leeuwenhoek. 2004 Aug;86(2):111-9. doi: 10.1023/B:ANTO.0000036118.41484.e7. Antonie Van Leeuwenhoek. 2004. PMID: 15280645 - Protein engineering of the milk-clotting aspartic proteinases.
Aikawa J, Nishiyama M, Beppu T. Aikawa J, et al. Scand J Clin Lab Invest Suppl. 1992;210:51-8. Scand J Clin Lab Invest Suppl. 1992. PMID: 1455180 Review. - Substrate specificity and inhibitors of aspartic proteinases.
Kay J, Dunn BM. Kay J, et al. Scand J Clin Lab Invest Suppl. 1992;210:23-30. Scand J Clin Lab Invest Suppl. 1992. PMID: 1455176 Review.
Cited by
- Rhizomucor miehei triglyceride lipase is processed and secreted from transformed Aspergillus oryzae.
Huge-Jensen B, Andreasen F, Christensen T, Christensen M, Thim L, Boel E. Huge-Jensen B, et al. Lipids. 1989 Sep;24(9):781-5. doi: 10.1007/BF02544584. Lipids. 1989. PMID: 2586234 - Rhizomucor miehei triglyceride lipase is synthesized as a precursor.
Boel E, Huge-Jensen B, Christensen M, Thim L, Fiil NP. Boel E, et al. Lipids. 1988 Jul;23(7):701-6. doi: 10.1007/BF02535672. Lipids. 1988. PMID: 3419283 - Thermophilic fungi: their physiology and enzymes.
Maheshwari R, Bharadwaj G, Bhat MK. Maheshwari R, et al. Microbiol Mol Biol Rev. 2000 Sep;64(3):461-88. doi: 10.1128/MMBR.64.3.461-488.2000. Microbiol Mol Biol Rev. 2000. PMID: 10974122 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources