The pattern of MAP-2 binding on microtubules: visual enhancement of MAP attachment sites by antibody labeling and electron microscopy

Roberta A. Gottlieb, Douglas B. Murphy

Research output: Contribution to journalArticle

Abstract

We used affinity-purified rabbit antibody to hog brain microtubule-associated protein 2 (MAP-2) to examine the pattern of attachment of MAPs to microtubules purified by cycles of in vitro assembly and disassembly. Microtubules were fixed, deposited on EM grids, and labeled with antibody and protein A-gold colloid followed by negative staining. We observed that: (1) The sites of MAP attachment were greatly enhanced by antibody binding in negatively stained preparations. (2) The axial repeat revealed by antibody (100 ± 5 nm) was greater than the previously reported value of 32 nm based on thin sectioning and negative staining procedures. (3) The antibody was arranged in a broad band and revealed a helical pattern of binding. (4) Microtubules with and without treatment with α-chymotrypsin to remove the projection portion of MAP-2 looked similar, suggesting that the antibody-enhanced pattern may reflect the sites of MAP attachment on microtubules. (5) Microtubules with an increased MAP:tubulin ratio exhibited the same 100-nm periodicity.

Original languageEnglish (US)
Pages (from-to)175-185
Number of pages11
JournalJournal of Ultrasructure Research
Volume85
Issue number2
DOIs
StatePublished - 1983

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Microtubule-Associated Proteins
Protein Binding
Microtubules
Electron Microscopy
Antibodies
Negative Staining
Microtomy
Gold Colloid
Staphylococcal Protein A
Chymotrypsin
Periodicity
Tubulin
Rabbits
Brain

ASJC Scopus subject areas

  • Molecular Biology
  • Anatomy

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The pattern of MAP-2 binding on microtubules : visual enhancement of MAP attachment sites by antibody labeling and electron microscopy. / Gottlieb, Roberta A.; Murphy, Douglas B.

In: Journal of Ultrasructure Research, Vol. 85, No. 2, 1983, p. 175-185.

Research output: Contribution to journalArticle

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