| Microtubule minus ends can be labelled with a phage display antibody specific to alpha-tubulin |
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| Author(s): Fan J, Griffiths AD, Lockhart A, Cross RA, Amos LA |
| Source: JOURNAL OF MOLECULAR BIOLOGY Volume: 259 Issue: 3 Pages: 325-330 Published: JUN 14 1996 |
| Times Cited: 43 References: 19 |
| Abstract: To investigate the orientation of alpha- and beta-tubulin heterodimers within microtubules, we cloned a phage display antibody to alpha-tubulin. The N-terminal 100 residues of alpha-tubulin were bacterially expressed and used to select clones from a large repertoire of antibody-expressing phagemid particles. One clone reacted with the expressed alpha-tubulin N terminus and native tubulin dimer but not with the expressed beta-tubulin N terminus. Electron microscopy showed 30 nm gold beads coated with die antibody binding to one end of brain microtubules. The beads bound to the minus ends of axonemes but not to the brain tubulin extensions from their plus ends. In sliding motility assays with a plus end directed motor, beads were pushed ahead of the microtubules. Our results indicate that an N-terminal epitope of alpha-tubulin is exposed only at the minus ends of microtubules. (C) 1996 Academic Press Limited |
| Document Type: Article |
| Language: English |
Addresses:
1. MRC, MOLEC BIOL LAB, CAMBRIDGE CB2 2QH, ENGLAND
2. MARIE CURIE INST, SURREY RH8 0TL, ENGLAND |
| Publisher: ACADEMIC PRESS LTD, 24-28 OVAL RD, LONDON, ENGLAND NW1 7DX |
| Subject Category: Biochemistry & Molecular Biology |
| IDS Number: UP667 |
| ISSN: 0022-2836 |