spacer gif spacer gif spacer gif spacer gif spacer gif
QUICK SEARCH:   [advanced]


spacer gif
     Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents    

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Linck, R. W.
Right arrow Articles by Steffen, W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Linck, R. W.
Right arrow Articles by Steffen, W.

Journal of Cell Science, Vol 88, Issue 4 453-466, Copyright © 1987 by Company of Biologists

JOURNAL ARTICLES

Characterization of antibodies as probes for structural and biochemical studies of tektins from ciliary and flagellar microtubules

RW Linck, MJ Goggin, JM Norrander and W Steffen
Department of Cell Biology and Neuroanatomy, University of Minnesota, Minneapolis 55455.

Rabbit antibodies raised and purified against three tektins, proteins of flagellar doublet microtubules from sea-urchin sperm (Lytechinus pictus and Strongylocentrotus purpuratus), were used to study tektin biochemistry and their structural localization. Doublet microtubules were fractionated into tektin filaments and separated by SDS-PAGE into three major tektin polypeptide bands (Mr = 47, 51 and 55 (X 10(3)), which were used to immunize rabbits. Antibodies against each tektin (anti-tektins) were affinity-purified and then characterized by two-dimensional isoelectric focusing/SDS-PAGE immunoblotting and by immunofluorescence microscopy. In two-dimensional immunoblots of 0.5% Sarkosyl-resistant fractions of flagellar microtubules, the antibody against the 55 X 10(3) Mr tektin (anti-55) stained one major polypeptide of 55 X 10(3) Mr and pI 6.9, anti-51 stained two polypeptides of 51 X 10(3) Mr and pI approximately 6.15, and anti-47 stained one major polypeptide of 47 X 10(3) Mr and pI 6.15. The anti-tektins also stained several minor neighbouring polypeptides, which may be isoelectric variants, novel tektins or unrelated proteins. Furthermore, anti-47 crossreacted with the major 55 X 10(3) Mr polypeptide. By immunofluorescence microscopy all three anti-tektins stained methanol-fixed echinoderm sperm flagella and embryonic cilia. In addition, anti-47 and anti-55 stained unfixed, demembranated axonemes. Besides staining axonemes, all anti-tektins labelled the basal body region, and anti-51 labelled the sperm head envelope. These results indicate that the tektins are a complex family of proteins that are components of axonemal microtubules and possibly other cytoplasmic and nuclear structures.


This article has been cited by other articles:


Home page
 Mol. Biol. CellHome page
J. M. Norrander, A. M. deCathelineau, J. A. Brown, M. E. Porter, and R. W. Linck
The Rib43a Protein Is Associated with Forming the Specialized Protofilament Ribbons of Flagellar Microtubules in Chlamydomonas
Mol. Biol. Cell, January 1, 2000; 11(1): 201 - 215.
[Abstract] [Full Text]

Home page
 J. Cell Sci.Home page
W Steffen, E. Fajer, and R. Linck
Centrosomal components immunologically related to tektins from ciliary and flagellar microtubules
J. Cell Sci., January 8, 1994; 107(8): 2095 - 2105.
[Abstract] [PDF]

Home page
 J. Cell Sci.Home page
R Chen, C. Perrone, L. Amos, and R. Linck
Tektin B1 from ciliary microtubules: primary structure as deduced from the cDNA sequence and comparison with tektin A1
J. Cell Sci., January 11, 1993; 106(3): 909 - 918.
[Abstract] [PDF]


© The Company of Biologists Ltd 1987