Microtubule-binding proteins from carrot - I. Initial characterization and microtubule bundling

Richard J. Cyr, Barry A. Palevitz

Research output: Contribution to journalArticle

88 Citations (Scopus)

Abstract

Microtubules (MTs) participate in several processes of fundamental importance to growth and development in higher plants, yet little is known about the proteins with which they associate. Information about these molecules is important because they probably play a role in mediating functional and structural differences between various MT arrays. As a first step in gaining insight into this problem, we have isolated, from suspension-cultured cells of carrot (Daucus carota L.), non-tubulin proteins which bind to and affect microtubules (MTs) in vitro. These proteins were isolated using taxol-stabilized neuronal MTs as an affinity substrate. They cause MT bundling at substoichiometric concentrations, support the assembly of tubulin in vitro, and at low concentrations, decorate single MTs in a periodic fashion. The bundled MTs formed in vitro share similarities with those seen in situ in a variety of plant cells, including a center-center spacing of 34 nm, cold stability, resistance to anti-microtubule drugs, and sensitivity to calcium. The bundling activity is specific; other cationic proteins, as well as poly-L-lysine, do not behave in a similar manner. The bundling activity is insensitive to ATP. By assaying bundling activity with dark-field microscopy and employing standard biochemical procedures, a small number of polypeptides involved in the bundling process were identified. Affinity-isolated antibodies to one of these polypeptides (Mr=76000) were found to co-localize with MTs in the cortical array of protoplasts. Our findings are discussed with reference to the importance of these proteins in the cell and to their relationship to microtubule-associated proteins in other eukaryotes.

Original languageEnglish (US)
Pages (from-to)245-260
Number of pages16
JournalPlanta
Volume177
Issue number2
DOIs
StatePublished - Feb 1 1989

Fingerprint

Microtubule Proteins
Daucus carota
Microtubules
carrots
microtubules
binding proteins
Carrier Proteins
proteins
Proteins
polypeptides
cold stability
Peptides
Antibody Affinity
Microtubule-Associated Proteins
Protoplasts
Plant Cells
paclitaxel
Tubulin
Paclitaxel
Eukaryota

All Science Journal Classification (ASJC) codes

  • Genetics
  • Plant Science

Cite this

Cyr, Richard J. ; Palevitz, Barry A. / Microtubule-binding proteins from carrot - I. Initial characterization and microtubule bundling. In: Planta. 1989 ; Vol. 177, No. 2. pp. 245-260.
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Microtubule-binding proteins from carrot - I. Initial characterization and microtubule bundling. / Cyr, Richard J.; Palevitz, Barry A.

In: Planta, Vol. 177, No. 2, 01.02.1989, p. 245-260.

Research output: Contribution to journalArticle

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