Immunopurification of a sarcomeric junctional protein complex containing GAPDH

Kevin A. McDonald, John Muschler, Alan F. Horwitz

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

We have isolated a monoclonal antibody, P4B2, which localizes to multiple anchorage junctions, namely, a subset of focal adhesions, the Z- disk of muscle, and neuromuseular junctions. Immunopurification of the antigen to this antibody from chicken brain tissue yielded a complex of three prominent proteins with mobilities of 36, 30, and 18 kDa. Amino acid sequencing of the purified proteins identified the 36-kDa protein as glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The other two protein bands were heterogeneous, containing proteins found in the synaptic vesicle fusion core complex. Immunolocalization of P4B2 antigen in developing cultured muscle cells showed that the antigen is incorporated into Z-lines soon after the sarcomeric architecture was positive for α-actinin. Together, the data indicate the P4B2 antigen is part of a unique GAPDH-containing protein complex that may be involved in reinforcement of established cytoskeletal structures.

Original languageEnglish (US)
Pages (from-to)305-318
Number of pages14
JournalExperimental Cell Research
Volume243
Issue number2
DOIs
StatePublished - Sep 15 1998
Externally publishedYes

Fingerprint

Glyceraldehyde-3-Phosphate Dehydrogenases
Antigens
Proteins
Actinin
Focal Adhesions
Synaptic Vesicles
Protein Sequence Analysis
Muscle Cells
Chickens
Cultured Cells
Monoclonal Antibodies
Muscles
Antibodies
Brain

Keywords

  • Focal adhesion
  • Glyceraldehyde-3-phosphate dehydrogenase
  • Integrin
  • Junctions
  • Z-disk

ASJC Scopus subject areas

  • Cell Biology

Cite this

Immunopurification of a sarcomeric junctional protein complex containing GAPDH. / McDonald, Kevin A.; Muschler, John; Horwitz, Alan F.

In: Experimental Cell Research, Vol. 243, No. 2, 15.09.1998, p. 305-318.

Research output: Contribution to journalArticle

McDonald, Kevin A. ; Muschler, John ; Horwitz, Alan F. / Immunopurification of a sarcomeric junctional protein complex containing GAPDH. In: Experimental Cell Research. 1998 ; Vol. 243, No. 2. pp. 305-318.
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