Antibodies that can discriminate between dynein heavy chains and their HUV1 photoproducts.
Dyneins are multi-subunit enzymes that transduce chemical energy into the mechanical energy that makes cilia and flagella beat and moves organelles towards the minus end of microtubules. The ATPase activity is borne by heavy chains, and recent molecular analysis indicates that dynein heavy chain genes form an ancient multigene family: the similarity between the same isoform of two distantly related species is greater than that between different isoforms of the same species. We have exploited sequence identities between a Paramecium axonemal dynein heavy chain gene cloned in our laboratory and sequences of dynein heavy chains from other species to prepare antibodies against active-site peptides capable of recognizing dynein heavy chains regardless of species or isoform. One of the antibodies is perfectly specific for the larger product of V1 photolysis (HUV1) and thus incorporates a unique property of the hydrolytic ATP binding site of all known dynein heavy chains, the capacity for photocleavage in the presence of micromolar vanadate. Our characterization of these reagents suggests that they will be useful for biochemical and in situ studies of known dyneins as well as identification of potential new members of the family.
Sperling
L
L
Centre de Génétique Moléculaire, CNRS, Gif-sur-Yvette, France.
Houari
A
A
Moudjou
M
M
Mazarguil
H
H
Wright
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology*; D000918:Antibody Specificity; D001665:Binding Sites; D002923:Cilia / Q000737:chemistry; D004398:Dyneins / Q000737:chemistry / Q000276:immunology* / Q000528:radiation effects*; D008969:Molecular Sequence Data; D016805:Paramecium tetraurelia / Q000235:genetics / Q000378:metabolism; D010782:Photolysis; D015800:Protozoan Proteins / Q000737:chemistry / Q000276:immunology* / Q000528:radiation effects*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000528:radiation effects; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014466:Ultraviolet Rays; D014638:Vanadates / Q000494:pharmacology
D000913:Antibodies, Protozoan; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D014638:Vanadates; D004398:Dyneins