The identification and characterisation of an actin-binding site in alpha-actinin by mutagenesis.
We have shown previously that the N-terminal actin-binding domain of alpha-actinin retains activity when expressed in E. coli as a fusion protein with glutathione-S-transferase. In the present study we have made a series of N- and C-terminal deletions within this domain and show that an actin-binding site is contained within residues 120-134. Amino acid substitutions within this region indicate that several highly conserved hydrophobic residues are involved in binding to F-actin. The hypothesis that the interaction between alpha-actinin and F-actin is predominantly hydrophobic in nature is supported by the observation that binding is relatively independent of salt concentration.
Kuhlman
P A
PA
Department of Biochemistry, University of Leicester, UK.
Hemmings
L
L
Critchley
D R
DR
D000185:Actinin / Q000235:genetics*; D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002645:Chickens / Q000235:genetics*; D004252:DNA Mutational Analysis; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics
D000199:Actins; D011993:Recombinant Fusion Proteins; D000185:Actinin; D005982:Glutathione Transferase