Actin-related proteins in Actinobacillus pleuropneumoniae and their interactions with actin-binding proteins.
A group of prokaryotic actin-related proteins (PARP) with an Mr of 43000 was detected in Actinobacillus pleuropneumoniae. These proteins were enriched by a depolymerization/polymerization cycle, under similar conditions to those used to polymerize muscle actin, and purified by affinity chromatography on a DNase I-Sepharose column. Three isoforms of A. pleuropneumoniae PARP (Ap-PARP) with pI values of 5.8, 6.15 and 6.2 were detected. Ap-PARP were recognized by four different anti-actin antibodies (one anti-muscle and three anti-cytoplasmic isoforms). Ap-PARP were also recognized by antibodies against Anabaena variabilis PARP (Av-PARP) and against actin-binding proteins such as alpha-actinin and spectrin, and also by a monoclonal antibody against heat-shock cognate protein 70 (Hsc70). Specific binding of phalloidin to Ap-PARP was detected both in permeabilized cells and in vitro. Purified Ap-PARP can polymerize under similar conditions to those required for skeletal muscle actin polymerization and the filaments formed appear to be decorated with myosin subfragment-1(S1) as observed by transmission electron microscopy. The amino acid composition of Ap-PARP revealed more similarities to muscle gamma-actin and the cytoplasmic beta-actin isoform than to eukaryotic actin-related proteins.
Guerrero-Barrera
Alma L
AL
Departamento de Biologı́a Celular, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 14-740, México, DF 07000, Mexico1.
de la Garza
Mireya
M
Departamento de Biologı́a Celular, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 14-740, México, DF 07000, Mexico1.
Mondragón
Ricardo
R
Departamento de Inmunologı́a, Escuela Nacional de Ciencias Biológicas IPN, Prolongación de Carpio y Plan de Ayala s/n, México, DF 11340, Mexico3.
Garcı A-Cuéllar
Claudia
C
División de Investigación Básica, Instituto Nacional de Cancerologı́a, San Fernando No 22, Tlalpan, DF 14000, Mexico4.|Departamento de Biologı́a Celular, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 14-740, México, DF 07000, Mexico1.
Segura-Nieto
Magdalena
M
Departamento de Ingenierı́a Genética de Plantas, Centro de Investigación y de Estudios Avanzados del IPN, Apartado Postal 629, Irapuato, Gto. 36500, Mexico2.
D016977:Actinobacillus pleuropneumoniae / Q000737:chemistry*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D015153:Blotting, Western; D002846:Chromatography, Affinity; D015638:Cytochalasin D / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D008840:Microfilament Proteins / Q000378:metabolism; D008854:Microscopy, Electron; D010590:Phalloidine / Q000378:metabolism; D011506:Proteins / Q000032:analysis / Q000502:physiology*; D013045:Species Specificity; D013050:Spectrometry, Fluorescence; D013552:Swine
D000596:Amino Acids; D008840:Microfilament Proteins; D011506:Proteins; D010590:Phalloidine; D015638:Cytochalasin D