Characterization of the cell surface heterodimer VLA-4 and related peptides

M. E. Hemler, C. Huang, Y. Takada, L. Schwarz, J. L. Strominger, M. L. Clabby

Research output: Contribution to journalArticle

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Abstract

A monoclonal antibody (B-5G10) was produced which specifically recognizes the M(r) 150,000/130,000 VLA-4 complex on the surface of human cells. Crosslinking studies indicated that the M(r) 150,000 α4 subunit of VLA-4 is in noncovalent 1:1 associated with the M(r) 130,000 VLA β subunit. In the absence of crosslinking, the VLA-4 α4β subunit complex was easily dissociated, especially in Nonidet P-40 detergent, or at elevated pH (above 8.0). Studies of dissociated subunits showed that B-5G10 recognize an epitope on the M(r) 150,000 α4 subunit of VLA-4, whereas the β subunit is immunologically identical to the M(r) 130,000 β subunit common to all VLA heterodimes. VLA-4 is widely distributed on hematopoietic cells, including thymocytes, peripheral blood lymphocytes, monocytes, activated T cells, T and B lymphoblastoid cell lines, and myeloid cell lines. However, VLA-4 is only weakly expressed on most adherent cell lines tested. Immunoprecipitates of VLA-4 often contain additional proteins of M(r) 80,000 and M(r) 70,000. These are probably derived from the M(r) 150,000 α4 subunit because: 1) they are both recognized by anti-α4 sera, but not anti-β sera; 2) the sum of their sizes is equal to the size of α4; 3) they are selectively coexpressed with α4 and not other VLA α subunits; 4) the M(r) 80,000 protein has an identical NH2-terminal sequence to α4; 5) like α4, the M(r) 70,000 and 80,000 peptides can variably associate with the VLA β subunits; and 6) trypsin appears to cleave the M(r) 150,000 α4 subunit into products of M(r) 70,000 and 80,000.

Original languageEnglish
Pages (from-to)11478-11485
Number of pages8
JournalJournal of Biological Chemistry
Volume262
Issue number24
Publication statusPublished - 1987
Externally publishedYes

Fingerprint

Integrin alpha4beta1
Peptides
Cells
Cell Line
Crosslinking
T-cells
Lymphocytes
Myeloid Cells
Thymocytes
Serum
Detergents
Trypsin
Epitopes
Monocytes
Proteins
Blood
Monoclonal Antibodies
T-Lymphocytes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Hemler, M. E., Huang, C., Takada, Y., Schwarz, L., Strominger, J. L., & Clabby, M. L. (1987). Characterization of the cell surface heterodimer VLA-4 and related peptides. Journal of Biological Chemistry, 262(24), 11478-11485.

Characterization of the cell surface heterodimer VLA-4 and related peptides. / Hemler, M. E.; Huang, C.; Takada, Y.; Schwarz, L.; Strominger, J. L.; Clabby, M. L.

In: Journal of Biological Chemistry, Vol. 262, No. 24, 1987, p. 11478-11485.

Research output: Contribution to journalArticle

Hemler, ME, Huang, C, Takada, Y, Schwarz, L, Strominger, JL & Clabby, ML 1987, 'Characterization of the cell surface heterodimer VLA-4 and related peptides', Journal of Biological Chemistry, vol. 262, no. 24, pp. 11478-11485.
Hemler ME, Huang C, Takada Y, Schwarz L, Strominger JL, Clabby ML. Characterization of the cell surface heterodimer VLA-4 and related peptides. Journal of Biological Chemistry. 1987;262(24):11478-11485.
Hemler, M. E. ; Huang, C. ; Takada, Y. ; Schwarz, L. ; Strominger, J. L. ; Clabby, M. L. / Characterization of the cell surface heterodimer VLA-4 and related peptides. In: Journal of Biological Chemistry. 1987 ; Vol. 262, No. 24. pp. 11478-11485.
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abstract = "A monoclonal antibody (B-5G10) was produced which specifically recognizes the M(r) 150,000/130,000 VLA-4 complex on the surface of human cells. Crosslinking studies indicated that the M(r) 150,000 α4 subunit of VLA-4 is in noncovalent 1:1 associated with the M(r) 130,000 VLA β subunit. In the absence of crosslinking, the VLA-4 α4β subunit complex was easily dissociated, especially in Nonidet P-40 detergent, or at elevated pH (above 8.0). Studies of dissociated subunits showed that B-5G10 recognize an epitope on the M(r) 150,000 α4 subunit of VLA-4, whereas the β subunit is immunologically identical to the M(r) 130,000 β subunit common to all VLA heterodimes. VLA-4 is widely distributed on hematopoietic cells, including thymocytes, peripheral blood lymphocytes, monocytes, activated T cells, T and B lymphoblastoid cell lines, and myeloid cell lines. However, VLA-4 is only weakly expressed on most adherent cell lines tested. Immunoprecipitates of VLA-4 often contain additional proteins of M(r) 80,000 and M(r) 70,000. These are probably derived from the M(r) 150,000 α4 subunit because: 1) they are both recognized by anti-α4 sera, but not anti-β sera; 2) the sum of their sizes is equal to the size of α4; 3) they are selectively coexpressed with α4 and not other VLA α subunits; 4) the M(r) 80,000 protein has an identical NH2-terminal sequence to α4; 5) like α4, the M(r) 70,000 and 80,000 peptides can variably associate with the VLA β subunits; and 6) trypsin appears to cleave the M(r) 150,000 α4 subunit into products of M(r) 70,000 and 80,000.",
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AB - A monoclonal antibody (B-5G10) was produced which specifically recognizes the M(r) 150,000/130,000 VLA-4 complex on the surface of human cells. Crosslinking studies indicated that the M(r) 150,000 α4 subunit of VLA-4 is in noncovalent 1:1 associated with the M(r) 130,000 VLA β subunit. In the absence of crosslinking, the VLA-4 α4β subunit complex was easily dissociated, especially in Nonidet P-40 detergent, or at elevated pH (above 8.0). Studies of dissociated subunits showed that B-5G10 recognize an epitope on the M(r) 150,000 α4 subunit of VLA-4, whereas the β subunit is immunologically identical to the M(r) 130,000 β subunit common to all VLA heterodimes. VLA-4 is widely distributed on hematopoietic cells, including thymocytes, peripheral blood lymphocytes, monocytes, activated T cells, T and B lymphoblastoid cell lines, and myeloid cell lines. However, VLA-4 is only weakly expressed on most adherent cell lines tested. Immunoprecipitates of VLA-4 often contain additional proteins of M(r) 80,000 and M(r) 70,000. These are probably derived from the M(r) 150,000 α4 subunit because: 1) they are both recognized by anti-α4 sera, but not anti-β sera; 2) the sum of their sizes is equal to the size of α4; 3) they are selectively coexpressed with α4 and not other VLA α subunits; 4) the M(r) 80,000 protein has an identical NH2-terminal sequence to α4; 5) like α4, the M(r) 70,000 and 80,000 peptides can variably associate with the VLA β subunits; and 6) trypsin appears to cleave the M(r) 150,000 α4 subunit into products of M(r) 70,000 and 80,000.

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