Preliminary structure analysis of the DH/PH domains of leukemia-associated RhoGEF.
Leukemia-associated RhoGEF (LARG) is a multidomain protein that relays signals from Galpha(12/13)-coupled heptahelical receptors to GTPases that regulate the cytoskeleton. To understand the molecular basis of LARG-mediated signal transduction, structural analysis of its DH/PH domains has been initiated. The LARG DH/PH domains have been overexpressed in Escherichia coli as a TEV protease-cleavable fusion protein containing maltose-binding protein and a hexahistidine tag at the N- and C-termini, respectively. Crystals of the DH/PH domains were obtained (space group C2; unit-cell parameters a = 195.5, b = 46.0, c = 75.1 A, beta = 105.0 degrees ) and xenon and NaBr derivatives were generated which should allow the structure to be determined by MIRAS.
D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020662:Guanine Nucleotide Exchange Factors / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D064067:Rho Guanine Nucleotide Exchange Factors; D017386:Sequence Homology, Amino Acid
C577222:ARHGEF12 protein, human; D020662:Guanine Nucleotide Exchange Factors; D011993:Recombinant Fusion Proteins; D064067:Rho Guanine Nucleotide Exchange Factors