Purification and functional analyses of ALS2 and its homologue.
ALS2 is a causative gene product for a form of the familial motor neuron diseases. Computational genomic analysis identified ALS2CL, which is a novel protein highly homologous to the C-terminal region of ALS2. Both proteins contain the VPS9 domain, which is a hallmark for all known members of the guanine nucleotide exchange factors for Rab5 (Rab5GEF), and are known to act as novel factors modulating the Rab5-mediated endosome dynamics in the cells. It has also been reported that oligomerization of ALS2 is one of the fundamental features of its biochemical and physiological function involving endosome dynamics. This chapter describes methods, including purification of the recombinant ALS2 and ALS2CL, and Rab5GEF assay, which have been utilized to clarify the molecular function for ALS2 and ALS2CL.
Shinji
S
Hadano
Joh-E
JE
Ikeda
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002352:Carrier Proteins / Q000302:isolation & purification* / Q000378:metabolism; D002522:Chlorocebus aethiops; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D020662:Guanine Nucleotide Exchange Factors; D006801:Humans; D011485:Protein Binding; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D020696:rab5 GTP-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism
C493234:ALS2CL protein, human; D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D017931:DNA Primers; D020662:Guanine Nucleotide Exchange Factors; D011994:Recombinant Proteins; D020696:rab5 GTP-Binding Proteins