1T1I

High Resolution Crystal Structure of Mutant W129A of Kumamolisin, a Sedolisin Type Proteinase (previously called Kumamolysin or KSCP)

Summary for 1T1I

• Entry DOI: 10.2210/pdb1t1i/pdb
• Related: 1GT9 1GTG 1GTJ 1GTL 1T1E 1T1G
• Descriptor: kumamolisin, CALCIUM ION, SULFATE ION, ... (4 entities in total)
• Functional Keywords: serine-carboxyl proteinases, subtilases, catalytic mechanism, sedolisin, kumamolisin, hydrolase
• Biological source: Bacillus sp. MN-32
• Total number of polymer chains: 1
• Total formula weight: 37103.77

Authors

Comellas-Bigler, M.,Maskos, K.,Huber, R.,Oyama, H.,Oda, K.,Bode, W. (deposition date: 2004-04-16, release date: 2004-08-03, Last modification date: 2024-02-14)

Primary citation

Comellas-Bigler, M.,Maskos, K.,Huber, R.,Oyama, H.,Oda, K.,Bode, W.
1.2 a crystal structure of the serine carboxyl proteinase pro-kumamolisin: structure of an intact pro-subtilase
Structure, 12:1313-1323, 2004

PubMed Abstract: Kumamolisin, an extracellular proteinase derived from an acido/thermophilic Bacillus, belongs to the sedolisin family of endopeptidases characterized by a subtilisin-like fold and a Ser-Glu-Asp catalytic triad. In kumamolisin, the Asp82 carboxylate hydrogen bonds to Glu32-Trp129, which might act as a proton sink stabilizing the catalytic residues. The 1.2/1.3 A crystal structures of the Glu32-->Ala and Trp129-->Ala mutants show that both mutations affect the active-site conformation, causing a 95% activity decrease. In addition, the 1.2 A crystal structure of the Ser278-->Ala mutant of pro-kumamolisin was determined. The prodomain exhibits a half-beta sandwich core docking to the catalytic domain similarly as the equivalent subtilisin prodomains in their catalytic-domain complexes. This pro-kumamolisin structure displays, for the first time, the uncleaved linker segment running across the active site and connecting the prodomain with the properly folded catalytic domain. The structure strongly points to an initial intramolecular activation cleavage in subtilases, as presumed for pro-subtilisin and pro-furin.

PubMed: 15242607
DOI: 10.1016/j.str.2004.04.013

Experimental method

X-RAY DIFFRACTION (1.28 Å)