Biochemical characterization of Alsin, a Rab5 and Rac1 guanine nucleotide exchange factor

Justin D. Topp, Darren S. Carney, Bruce F Horazdovsky

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Alsin is the gene product mutated in three juvenile-onset neurodegenerative disorders including amyotrophic lateral sclerosis 2 (ALS2). Sequence motif searches within Alsin predict the presence of Vps9, DH, and PH domains, implying that Alsin may function as a guanine nucleotide exchange factor (GEF) for Rab5 and a member of the Rho GTPase family. Procedures are presented in this chapter for the expression, purification, and biochemical characterization of the individual GEF domains of Alsin. A fractionation method is also described for the determination of Alsin's subcellular distribution. The presence of both Rac1 and Rab5 GEF activities makes Alsin a unique dual exchange factor that may couple endocytosis (via Rab5 activation) to cytoskeletal modulation (via Rac1 activation).

Original languageEnglish (US)
Article number22
Pages (from-to)261-276
Number of pages16
JournalMethods in Enzymology
Volume403
DOIs
StatePublished - 2005

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Guanine Nucleotide Exchange Factors
Chemical activation
rho GTP-Binding Proteins
Amyotrophic Lateral Sclerosis
Fractionation
Endocytosis
Neurodegenerative Diseases
Purification
Genes
Modulation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Biochemical characterization of Alsin, a Rab5 and Rac1 guanine nucleotide exchange factor. / Topp, Justin D.; Carney, Darren S.; Horazdovsky, Bruce F.

In: Methods in Enzymology, Vol. 403, 22, 2005, p. 261-276.

Research output: Contribution to journalArticle

Topp, Justin D. ; Carney, Darren S. ; Horazdovsky, Bruce F. / Biochemical characterization of Alsin, a Rab5 and Rac1 guanine nucleotide exchange factor. In: Methods in Enzymology. 2005 ; Vol. 403. pp. 261-276.
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