Nondenaturing electrophoresis as a tool to investigate tubulin complexes.
A protein molecule may exist as a monomer, homo-oligomer, or hetero-oligomer in a multiprotein complex. One-dimensional (1-D) native electrophoresis has long been used to characterize tubulins and their complexes. In this chapter, we describe the simplest way to identify the state of aggregation of commercial or homemade tubulins for further studies based on 1-D electrophoresis under nondenaturing conditions. We present a series of detailed protocols that can be used to analyze the maturation of alpha- and beta-tubulins and to identify the complexes formed during the folding and dimerization pathway as well as their stability.
Fanarraga
Mónica López
ML
Departamento de Biología Molecular, Facultad de Medicina, IFIMAV-Universidad de Cantabria, 39011 Santander, Spain.
Carranza
Gerardo
G
Castaño
Raquel
R
Nolasco
Sofia
S
Avila
J
J
Zabala
J C
JC
D000818:Animals; D004586:Electrophoresis / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D006801:Humans; D008954:Models, Biological; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D055503:Protein Multimerization / Q000502:physiology; D055550:Protein Stability; D040901:Proteomics / Q000379:methods; D014404:Tubulin / Q000138:chemical synthesis / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*
D046912:Multiprotein Complexes; D014404:Tubulin