title: Purification, crystallization and X-ray crystallographic analysis of human RAB11(S20V), a constitutively active GTP-binding form.
abstract: 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 I4, 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).
authors.affiliation: Department of Biochemistry, Yeungnam University, Gyeongsan, Republic of Korea.
authors.forename: Chang Min
authors.initials: CM
authors.lastname: Kim
authors.affiliation: Department of Biochemistry, Yeungnam University, Gyeongsan, Republic of Korea.
authors.forename: Jae Young
authors.initials: JY
authors.lastname: Choi
authors.affiliation: Department of Biochemistry, Yeungnam University, Gyeongsan, Republic of Korea.
authors.forename: Jong Hwan
authors.initials: JH
authors.lastname: Yoon
authors.affiliation: Department of Biochemistry, Yeungnam University, Gyeongsan, Republic of Korea.
authors.forename: Hyun Ho
authors.initials: HH
authors.lastname: Park
mesh_terms: D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray; D006160:Guanosine Triphosphate / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D020691:rab GTP-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D000099193:rab11 GTP-Binding Proteins
chemical_list: D006160:Guanosine Triphosphate; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins
keywords: RAB11; crystallization; diffraction; membrane trafficking; small G protein