Strong association of bovine IgM with microvilli and their microfilament core from 13762 ascites tumor cells

Xuehai Ye, Thomas N. Metcalf, David Andrews, Coralie A Carothers Carraway, Kermit L. Carraway

Research output: Contribution to journalArticle

Abstract

Microfilament cores, obtained by extracting 13762 mammary ascites tumor cell microvilli with Triton X-100, contain a major glycoprotein migrating at an apparent molecular weight of 80 kDa by dodecyl sulfate-polyacrylamide gel electrophoresis. The 80-kDa component is a disulfide-linked multimer, as demonstrated by velocity sedimentation and agarose-acrylamide gel electrophoresis analyses under nonreducing conditions. This 80-kDa species is not metabolically labeled, as is a minor 80-kDa glycoprotein found in the cores, membranes, and an isolated transmembrane complex with actin. Antibodies prepared against the 80-kDa glycoprotein react strongly with bovine IgM and more weakly with rat IgM. These antibodies were used to demonstrate that the 80-kDa component is present in microvilli, microvillar microfilament cores, and microvillar membranes only if the microvilli are prepared in the presence of calf serum. The 80-kDa component, purified by velocity sedimentation in dodecyl sulfate, reacts with anti-rat IgM by immunoblot analyses. Moreover, immunoprecipitation of detergent extracts of microvilli with anti-rat IgM specifically sediments the 80-kDa component. The 80-kDa glycoprotein fractionates with the actin-containing transmembrane complex prepared by gel filtration of Triton-solubilized microvillar membranes. These results indicate that the disulfide-linked, multimeric 80-kDa component is bovine IgM, which binds strongly to a cell-surface component of the microvilli, and is indirectly associated with the microfilament cores. Thus, the IgM provides a marker by which the transmembrane complexes to the microfilaments can be identified.

Original languageEnglish
Pages (from-to)160-172
Number of pages13
JournalExperimental Cell Research
Volume182
Issue number1
DOIs
StatePublished - Jan 1 1989

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Microvilli
Actin Cytoskeleton
Ascites
Immunoglobulin M
Glycoproteins
Neoplasms
Disulfides
Membranes
Actins
Agar Gel Electrophoresis
Acrylamide
Antibodies
Octoxynol
Cellular Structures
Immunoprecipitation
Detergents
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Molecular Weight
Breast Neoplasms

ASJC Scopus subject areas

  • Cell Biology

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Strong association of bovine IgM with microvilli and their microfilament core from 13762 ascites tumor cells. / Ye, Xuehai; Metcalf, Thomas N.; Andrews, David; Carraway, Coralie A Carothers; Carraway, Kermit L.

In: Experimental Cell Research, Vol. 182, No. 1, 01.01.1989, p. 160-172.

Research output: Contribution to journalArticle

Ye, Xuehai ; Metcalf, Thomas N. ; Andrews, David ; Carraway, Coralie A Carothers ; Carraway, Kermit L. / Strong association of bovine IgM with microvilli and their microfilament core from 13762 ascites tumor cells. In: Experimental Cell Research. 1989 ; Vol. 182, No. 1. pp. 160-172.
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