RRC ID 3269
著者 Itoi Y, Horinaka M, Tsujimoto Y, Matsui H, Watanabe K.
タイトル Characteristic features in the structure and collagen-binding ability of a thermophilic collagenolytic protease from the thermophile Geobacillus collagenovorans MO-1.
ジャーナル J. Bacteriol.
Abstract A collagen-degrading thermophile, Geobacillus collagenovorans MO-1, extracellularly produces a collagenolytic protease with a large molecular mass. Complete nucleotide sequencing of this gene after gene cloning revealed that the collagenolytic protease is a member of the subtilisin family of serine proteases and consists of a signal sequence for secretion, a prosequence for maturation, a catalytic region, 14 direct repeats of 20 amino acids at the C terminus, and a region with unknown function intervening between the catalytic region and the numerous repeats. Since the unusual repeats are most likely to be cleaved in the secreted form of the enzyme, the intervening region was investigated to determine whether it participates in collagen binding to facilitate collagen degradation. It was found that the mature collagenolytic protease containing the intervening region at the C terminus bound collagen but not the other insoluble proteins, elastin and keratin. Furthermore, the intervening region fused with glutathione S-transferase showed a collagen-binding ability comparable to that of the mature collagenolytic protease. The collagen-binding ability was finally attributed to two-thirds of the intervening region which is rich in beta-strands and is approximately 35 kDa in molecular mass. In the collagenolytic protease from strain MO-1, hydrogen bonds most likely predominate over the hydrophobic interaction for collagen binding, since a higher concentration of NaCl released collagen from the enzyme surface but a nonionic detergent could not. To the best of our knowledge, this is the first report of a thermophilic collagenolytic protease containing the collagen-binding segment.
巻・号 188(18)
ページ 6572-9
公開日 2006-9-6
DOI 10.1128/JB.00767-06
PII 188/18/6572
PMID 16952949
PMC PMC1595469
MeSH Amino Acid Sequence Bacillaceae / enzymology* Base Sequence Catalytic Domain Cloning, Molecular Collagen / metabolism* DNA, Bacterial / chemistry DNA, Bacterial / genetics Elastin / metabolism Keratins / metabolism Molecular Sequence Data Molecular Weight Protein Binding Protein Interaction Mapping Protein Sorting Signals Repetitive Sequences, Amino Acid / genetics Sequence Analysis, DNA Sequence Homology, Amino Acid Serine Endopeptidases / chemistry Serine Endopeptidases / genetics* Serine Endopeptidases / metabolism*
IF 3.143
引用数 26
原核生物(大腸菌) CV-25(DH5alpha)?