title: Crystallization and preliminary X-ray diffraction analysis of an active-site mutant of `loopless' family GH19 chitinase from Bryum coronatum in a complex with chitotetraose.
abstract: The catalytic mechanism of family GH19 chitinases is not well understood owing to insufficient information regarding the three-dimensional structures of enzyme-substrate complexes. Here, the crystallization and preliminary X-ray diffraction analysis of a selenomethionine-labelled active-site mutant of `loopless' family GH19 chitinase from the moss Bryum coronatum in complex with chitotetraose, (GlcNAc)4, are reported. The crystals were grown using the vapour-diffusion method. They diffracted to 1.58 Å resolution using synchrotron radiation at the Photon Factory. The crystals belonged to the monoclinic space group C2, with unit-cell parameters a = 74.5, b = 58.4, c = 48.1 Å, β = 115.6°. The asymmetric unit of the crystals is expected to contain one protein molecule, with a Matthews coefficient of 2.08 Å(3) Da(-1) and a solvent content of 41%.
authors: Department of Advanced Bioscience, Kinki University, 3327-204 Nakamachi, Nara 631-8505, Japan.
authors: Takayuki
authors: T
authors: Ohnuma
authors: Naoyuki
authors: N
authors: Umemoto
authors: Toki
authors: T
authors: Taira
authors: Tamo
authors: T
authors: Fukamizo
authors: Tomoyuki
authors: T
authors: Numata
mesh_terms: D044002:Bryophyta / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D002688:Chitinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017356:Synchrotrons
chemical_list: D009844:Oligosaccharides; D010940:Plant Proteins; D011994:Recombinant Proteins; C012238:chitotetrose; D002688:Chitinases
keywords: Bryum coronatum; chitotetraose; co-crystallization; loopless GH19 chitinase