期刊
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY
卷 71, 期 -, 页码 565-577出版社
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S1399004714027722
关键词
polycystic kidney disease-like domains; ColG; ColH; Clostridium histolyticum
资金
- National Institutes of Health [GM103429, GM103450]
- Arkansas Biosciences Institute
- Japan Society for the Promotion of Science
- Kagawa University
- NSF
- Honors College of UA
- Arkansas Department of Higher Education
- Doctoral Academy Fellowship
- Department of Energy
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [P30GM103450, P20GM103429] Funding Source: NIH RePORTER
Clostridium histolyticum collagenases ColG and ColH are segmental enzymes that are thought to be activated by Ca2+-triggered domain reorientation to cause extensive tissue destruction. The collagenases consist of a collagenase module (s1), a variable number of polycystic kidney disease-like (PKD-like) domains (s2a and s2b in ColH and s2 in ColG) and a variable number of collagen-binding domains (s3 in ColH and s3a and s3b in ColG). The X-ray crystal structures of Ca2+-bound holo s2b (1.4 angstrom resolution, R = 15.0%, R-free = 19.1%) and holo s2a (1.9 angstrom resolution, R = 16.3%, R-free = 20.7%), as well as of Ca2+-free apo s2a (1.8 angstrom resolution, R = 20.7%, R-free = 27.2%) and two new forms of N-terminally truncated apo s2 (1.4 angstrom resolution, R = 16.9%, R-free = 21.2%; 1.6 angstrom resolution, R = 16.2%, R-free = 19.2%), are reported. The structurally similar PKD-like domains resemble the V-set Ig fold. In addition to a conserved beta-bulge, the PKD-like domains feature a second bulge that also changes the allegiance of the subsequent beta-strand. This beta-bulge and the genesis of a Ca2+ pocket in the archaeal PKD-like domain suggest a close kinship between bacterial and archaeal PKD-like domains. Different surface properties and indications of different dynamics suggest unique roles for the PKD-like domains in ColG and in ColH. Surface aromatic residues found on ColH s2a-s2b, but not on ColG s2, may provide the weak interaction in the biphasic collagen-binding mode previously found in s2b-s3. B-factor analyses suggest that in the presence of Ca2+ the midsection of s2 becomes more flexible but the midsections of s2a and s2b stay rigid. The different surface properties and dynamics of the domains suggest that the PKD-like domains of M9B bacterial collagenase can be grouped into either a ColG subset or a ColH subset. The conserved properties of PKD-like domains in ColG and in ColH include Ca2+ binding. Conserved residues not only interact with Ca2+, but also position the Ca2+-interacting water molecule. Ca2+ aligns the N-terminal linker approximately parallel to the major axis of the domain. Ca2+ binding also increases stability against heat and guanidine hydrochloride, and may improve the longevity in the extracellular matrix. The results of this study will further assist in developing collagen-targeting vehicles for various signal molecules.
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