Immunopurification of a sarcomeric junctional protein complex containing GAPDH.
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 neuromuscular 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 alpha-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.
McDonald
K A
KA
Department of Biochemistry, University of Illinois, Urbana, Illinois 61801, USA. kmcdonal@uiuc.edu
Muschler
J
J
Horwitz
A F
AF
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000941:Antigens / Q000378:metabolism; D002645:Chickens; D003599:Cytoskeleton / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000302:isolation & purification / Q000378:metabolism*; D016023:Integrins / Q000378:metabolism; D007365:Intercellular Junctions / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009132:Muscles / Q000166:cytology; D011817:Rabbits; D012518:Sarcomeres / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000941:Antigens; D016023:Integrins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases