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Purification, crystallization and X‐ray crystallographic analysis of human RAB11(S20V), a constitutively active GTP‐binding form
Author(s) -
Kim Chang Min,
Choi Jae Young,
Yoon Jong Hwan,
Park Hyun Ho
Publication year - 2015
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x15015447
Subject(s) - gtp' , mutant , crystallography , chemistry , escherichia coli , endosome , crystallization , ras superfamily , angstrom , molecule , biochemistry , cell , enzyme , gene , organic chemistry
RAB11, a member of the Ras superfamily of small G proteins, is involved in the regulation of vesicle trafficking during endosome recycling. Substitution of Ser20 by Val20 in Rab11 [RAB11(S20V)] inhibits its GTP hydrolysis activity and produces a constitutively active GTP‐binding form. In this study, the RAB11(S20V) mutant was overexpressed in Escherichia coli with an engineered C‐terminal His tag. RAB11(S20V) was then purified to homogeneity and was crystallized at 293 K. X‐ray diffraction data were collected to a resolution of 2.4 Å from a crystal belonging to space group I 4, with unit‐cell parameters a = 74.11, b = 74.11, c = 149.44 Å. The asymmetric unit was estimated to contain two molecules of RAB11(S20V).