title: Immunochemical probing of the N-terminal segment on actin: the polymerization reaction.
abstract: The N-terminal segment of actin contains a cluster of acidic residues which are implicated in macromolecular interactions of this protein. In this work, the interrelationship between the N-terminal segment and the polymerization of actin was studied by using affinity-purified antibodies directed against the first seven N-terminal residues on alpha-skeletal actin (S alpha N). The Fab fragments of these antibodies showed equal affinities for G- and F-actin while the bivalent IgG bound preferentially to the polymerized actin. As monitored by pyrene fluorescence measurements, the binding of Fab to G-actin did not alter the kinetics of the MgCl2-induced polymerization; IgG accelerated this reaction considerably. Consistent with these observations, the binding of Fab to F-actin did not change its morphological appearance in electron micrographs and had no effect on the stability and the rate of dissociation of actin filaments. These results are discussed in terms of their implications to the spatial relationship between the N-terminal segment and the rest of the molecule and the context of the polymerization reaction of actin in vitro and in vivo.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, Los Angeles 90024.
authors.forename: G
authors.initials: G
authors.lastname: DasGupta
authors.forename: J
authors.initials: J
authors.lastname: White
authors.forename: M
authors.initials: M
authors.lastname: Phillips
authors.forename: J C
authors.initials: JC
authors.lastname: Bulinski
authors.forename: E
authors.initials: E
authors.lastname: Reisler
mesh_terms: D000199:Actins / Q000276:immunology / Q000648:ultrastructure; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D004797:Enzyme-Linked Immunosorbent Assay; D005453:Fluorescence; D015335:Molecular Probes; D008969:Molecular Sequence Data; D011108:Polymers; D011721:Pyrenes; D011817:Rabbits
chemical_list: D000199:Actins; D000906:Antibodies; D015335:Molecular Probes; D011108:Polymers; D011721:Pyrenes