ISSN:
0886-1544
Keywords:
dynein
;
heavy chains
;
flagella
;
cilia
;
outer arms
;
inner arms
;
polyclonal antibodies
;
affinity-purification
;
Life and Medical Sciences
;
Cell & Developmental Biology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Medicine
Notes:
While studying cAMP-dependent dynein α-heavy chain phosphorylation, we found previously [Stephens and Prior, 1992: J. Cell Sci. 103:999-1012] that high salt extraction of sperm flagella from the mussel Mytilus edulis or the clam Spisula solidissima removed most visible dynein arms, accompanied by an amount of Mg+2-ATPase that correlated with the mass of dynein α-and β-heavy chains removed. However, although almost devoid of ATPase activity, such extracted axonemes retained one third of the heavy chain mass as two sets of electrophoretically-distinct, vanadate-cleavable, non-phosphorylated proteins. To explore the nature of these dynein-like proteins, antibodies to the α- and β-heavy chains were blot affinity-purified from a rabbit antiserum raised against gradient-purified Spisula 18-20S flagellar outer arm dynein. Although able to recognize common epitopes of the opposite chain type, neither the α-nor the β-heavy chain antibody recognized the tightly-bound proteins in either species, proving that they are immunologically distinct. While the β-antibody recognized its heavy chain homolog in gill cilia, the α-antibody did not, demonstrating immunological distinction between flagellar and ciliary dynein α-heavy chains. Immunization of a mouse with nitrocellulose strips containing one of the two tightly-bound Spisula flagellar proteins produced an antiserum that cross-reacted with each tightly-bound protein in both species and also recognized α- and β-heavy chains. The anti-molluscan serum cross-reacted strongly with sea urchin sperm flagellar dynein B-, C-, and D-bands, considered to be inner arm components, but not with sea urchin outer arm α- or β-heavy chains. These data indicate that the electro-phoretically and immunologically distinct, tightly-bound proteins of molluscan flagella are inner arm dynein heavy chains. © 1995 Wiley-Liss, Inc.
Additional Material:
6 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/cm.970300404
