Maturation processing and characterization of streptopain

Chiu Yueh Chen, Shih Chi Luo, Chih Feng Kuo, Yee Shin Lin, Jiunn Jong Wu, Ming T. Lin, Ching Chuan Liu, Wen Yih Jeng, Woei Jer Chuang

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Streptopain is a cysteine protease expressed by Streptococcus pyogenes. To study the maturation mechanism of streptopain, wild-type and Q186N, C192S, H340R, N356D and W357A mutant proteins were expressed in Escherichia coli and purified to homogeneity. Proteolytic analyses showed that the maturation of prostreptococcal pyrogenic exotoxin B zymogen (pro-SPE B) involves eight intermediates with a combination of cis- and trans-processing. Based on the sequences of these intermediates, the substrate specificity of streptopain favors a hydrophobic residue at the P2 site. The relative autocatalytic rates of these mutants exhibited the order Q186N > W357A > N356D, C192S, H340R. Interestingly, the N356D mutant containing protease activity could not be converted into the 28-kDa form by autoprocessing. This observation suggested that Asn356 might involve the cis-processing of the propeptide. In addition, the maturation rates of pro-SPE B with trypsin and plasmin were 10- and 60-fold slower than that with active mature streptopain. These findings indicate that active mature streptopain likely plays the most important role in the maturation of pro-SPE B under physiological conditions.

Original languageEnglish
Pages (from-to)17336-17343
Number of pages8
JournalJournal of Biological Chemistry
Volume278
Issue number19
DOIs
Publication statusPublished - 2003 May 9

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streptopain
Exotoxins
Enzyme Precursors
Processing
Cysteine Proteases
Streptococcus pyogenes
Fibrinolysin
Mutant Proteins
Substrate Specificity
Trypsin
Escherichia coli
Peptide Hydrolases

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Chen, Chiu Yueh ; Luo, Shih Chi ; Kuo, Chih Feng ; Lin, Yee Shin ; Wu, Jiunn Jong ; Lin, Ming T. ; Liu, Ching Chuan ; Jeng, Wen Yih ; Chuang, Woei Jer. / Maturation processing and characterization of streptopain. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 19. pp. 17336-17343.
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Maturation processing and characterization of streptopain. / Chen, Chiu Yueh; Luo, Shih Chi; Kuo, Chih Feng; Lin, Yee Shin; Wu, Jiunn Jong; Lin, Ming T.; Liu, Ching Chuan; Jeng, Wen Yih; Chuang, Woei Jer.

In: Journal of Biological Chemistry, Vol. 278, No. 19, 09.05.2003, p. 17336-17343.

Research output: Contribution to journalArticle

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