Bovine platelets contain a 280 kDa microtubule-associated protein antigenically related to brain MAP 2

Fern Tablin, M. D. Castro

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Resting bovine platelets contain a microtubule coil which reorganizes into linear arrays upon thrombin activation. Microtubule arrays in both resting and activated platelets are extensively cross-linked. In an effort to determine the proteins responsible for this cross-linking, we have developed a method to isolate taxol-stabilized microtubule coils directly from platelet-rich plasma. Negatively stained coils are still cross-linked, and fine filamentous projections are seen between adjacent microtubules. Critical-point-dried rotary shadowed replicas of these coils most clearly demonstrate the projections radiating from individual microtubules as well as along the microtubule coil. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE) analysis of isolated coils shows many microtubule-associated proteins (MAPs) present in addition to tubulin. One of these proteins, a 280 kDa MAP, cross-reacts with an antibody to bovine brain MAP 2 by immunoblot analysis. Immunofluorescence localization of this protein with both monoclonal and polyclonal antibodies demonstrates that it is associated with the microtubule coil in resting platelets and with the linear microtubule array present after thrombin activation. Immunoelectron microscopic localization demonstrates that projections from individual microtubules are labeled by the antibodies. We suggest that this MAP, along with several other potential MAPs, is responsible for the cross-linking and stability of bovine platelet microtubules.

Original languageEnglish (US)
Pages (from-to)415-421
Number of pages7
JournalEuropean Journal of Cell Biology
Volume56
Issue number2
StatePublished - 1991

Fingerprint

Microtubule-Associated Proteins
Microtubules
Blood Platelets
Brain
Thrombin
Proteins
Platelet-Rich Plasma
Antibodies
Tubulin
Paclitaxel
Sodium Dodecyl Sulfate
Fluorescent Antibody Technique
Polyacrylamide Gel Electrophoresis
Monoclonal Antibodies

Keywords

  • MAPs
  • Marginal bands
  • Microtubules
  • Nocodazole
  • Taxol

ASJC Scopus subject areas

  • Cell Biology
  • Anatomy

Cite this

Bovine platelets contain a 280 kDa microtubule-associated protein antigenically related to brain MAP 2. / Tablin, Fern; Castro, M. D.

In: European Journal of Cell Biology, Vol. 56, No. 2, 1991, p. 415-421.

Research output: Contribution to journalArticle

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AB - Resting bovine platelets contain a microtubule coil which reorganizes into linear arrays upon thrombin activation. Microtubule arrays in both resting and activated platelets are extensively cross-linked. In an effort to determine the proteins responsible for this cross-linking, we have developed a method to isolate taxol-stabilized microtubule coils directly from platelet-rich plasma. Negatively stained coils are still cross-linked, and fine filamentous projections are seen between adjacent microtubules. Critical-point-dried rotary shadowed replicas of these coils most clearly demonstrate the projections radiating from individual microtubules as well as along the microtubule coil. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE) analysis of isolated coils shows many microtubule-associated proteins (MAPs) present in addition to tubulin. One of these proteins, a 280 kDa MAP, cross-reacts with an antibody to bovine brain MAP 2 by immunoblot analysis. Immunofluorescence localization of this protein with both monoclonal and polyclonal antibodies demonstrates that it is associated with the microtubule coil in resting platelets and with the linear microtubule array present after thrombin activation. Immunoelectron microscopic localization demonstrates that projections from individual microtubules are labeled by the antibodies. We suggest that this MAP, along with several other potential MAPs, is responsible for the cross-linking and stability of bovine platelet microtubules.

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