Identification of tubulin isoforms in different tissues of Ascaris suum using anti-tubulin monoclonal antibodies.
Three monoclonal antibodies specific to alpha- and beta-tubulin were used to examine the expression of tubulin isoforms in the intestine, reproductive tract and body wall muscle of A. suum. The tubulins were found to be different in their isoelectric points, number of isoforms and peptide maps with Western blot analysis of one-dimensional polyacrylamide gel confirming the presence of alpha-, beta 1- and beta 2-tubulin. Commercial cross-reactive anti-alpha and anti-beta MAbs 356 and 357 recognized tubulin from A. suum tissues as well as from pig brain, whereas anti-A. suum beta-tubulin specific MAb P3D6 recognized tubulin from the A. suum tissues only. Two-dimensional gel analysis showed different isoform patterns in different A. suum tissues with anti-A. suum beta-tubulin MAb P3D6 and cross-reactive beta-tubulin MAb 357 recognizing 2-4 beta-tubulin isoforms and anti-alpha-tubulin MAb 356 recognizing 1-6 alpha-tubulin isoforms. Different peptide maps of tubulin were observed in the three tissues, when subjected to limited proteolysis followed by SDS-PAGE. The data indicate that different tubulins are found in different tissues of adult A. suum.
Bughio
N I
NI
Institute of Parasitology, McGill University, Macdonald College, Ste-Anne de Bellevue, Québec, Canada.
Faubert
G M
GM
Prichard
R K
RK
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001200:Ascaris / Q000737:chemistry*; D015153:Blotting, Western; D010449:Peptide Mapping; D014404:Tubulin / Q000032:analysis* / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D014404:Tubulin