Cloning of the cDNA encoding human nonmuscle myosin heavy chain-B and analysis of human tissues with isoform-specific antibodies

Carrie Phillips, Katsutoshi Yamakawa, Robert S. Adelstein

Research output: Contribution to journalArticle

103 Citations (Scopus)

Abstract

Previously, we reported the sequence of cDNA clones encoding amino acids 63 through 723 of the human nonmuscle myosin heavy chain-B isoform. In this paper, we present the derived sequence of the remaining 1303 amino acids along with 5′ and 3′ untranslated sequences. We made use of the differences between the derived nonmuscle myosin heavy chain-A and-B amino acid sequences to raise isoform-specific antibodies. Immunoblot analysis reveals a differential expression of both myosin heavy chain isoforms in a variety of human adult and foetal tissues and cells. When extracts of human adult aorta were subjected to gel electrophoresis, two distinct Coomassie Blue-stained bands and a fused band were seen migrating at approximately 200 kDa. These bands can be detected with four different specific antibodies recognizing the two different smooth muscle myosin heavy chain isoforms (204 kDa and 200 kDa) and the two different nonmuscle myosin heavy chain isoforms (A and B). Using immunohistochemistry, we confirmed the presence of the four different isoforms in adult and foetal aortas.

Original languageEnglish (US)
Pages (from-to)379-389
Number of pages11
JournalJournal of Muscle Research and Cell Motility
Volume16
Issue number4
DOIs
StatePublished - Aug 1995
Externally publishedYes

Fingerprint

Cloning
Organism Cloning
Protein Isoforms
Myosin Heavy Chains
Complementary DNA
Tissue
Antibodies
Amino Acids
Aorta
Smooth Muscle Myosins
Electrophoresis
nonmuscle myosin type IIB heavy chain
Amino Acid Sequence
Fetus
Clone Cells
Gels
Immunohistochemistry

ASJC Scopus subject areas

  • Physiology
  • Endocrinology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Cloning of the cDNA encoding human nonmuscle myosin heavy chain-B and analysis of human tissues with isoform-specific antibodies. / Phillips, Carrie; Yamakawa, Katsutoshi; Adelstein, Robert S.

In: Journal of Muscle Research and Cell Motility, Vol. 16, No. 4, 08.1995, p. 379-389.

Research output: Contribution to journalArticle

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