Biochemical identity and characterization of the mouse interleukin-2 receptor β and γ(c) subunits

T. R. Malek, R. K. Furse, M. L. Fleming, A. J. Fadell, Y. W. He

Research output: Contribution to journalArticle

Abstract

Although the mouse IL-2 receptor (IL-2R) β and γ(c) subunits have been identified by molecular cloning, the biochemical identity of these subunits has not yet been established. In the present study, the mouse IL-2R was biochemically characterized from cell lines expressing normal and aberrant IL-2R. Using novel monoclonal antibodies specific for the β or γ(c) subunits, we established that the M(r) of the β chain is 90,000-100,000 and that of the γ(c) subunit is 75,000-80,000. Analysis of transfected EL4 cells that expressed α, γ(c), and truncated β subunits or mutant EL4 cells, which selectively lacked cell surface γ(c), revealed that no other material migrated to a position on SDS-PAGE characteristic of IL-2/IL-2Rβ and IL- 2/IL-2Rγ(c) cross-linked complexes, respectively. Thus, the β and γ(c) subunits appear to be the sole IL-2R constituents of these IL-2 cross-linked complexes. The IL-2/IL-2Rγ(c), but not the IL-2/IL-2Rβ, complex exhibited enhanced mobility after SDS-polyacrylamide gel electrophoresis under nonreducing conditions, suggesting a more compact structure for γ(c) as a result of intrachain disulfide bonds. The primary posttranslational modification of the mouse β and γ(c) subunits is N-linked glycosylation. These biochemical studies reconcile past uncertainties concerning the subunit composition of the mouse IL-2R and are consistent with a model of the IL-2R containing only three subunits.

Original languageEnglish
Pages (from-to)447-454
Number of pages8
JournalJournal of Interferon and Cytokine Research
Volume15
Issue number5
StatePublished - Jan 1 1995

Fingerprint

Interleukin-2 Receptors
Interleukin-2
Polyacrylamide Gel Electrophoresis
Molecular Cloning
Post Translational Protein Processing
Glycosylation
Disulfides
Uncertainty
Monoclonal Antibodies
Cell Line

ASJC Scopus subject areas

  • Cell Biology
  • Immunology
  • Virology

Cite this

Biochemical identity and characterization of the mouse interleukin-2 receptor β and γ(c) subunits. / Malek, T. R.; Furse, R. K.; Fleming, M. L.; Fadell, A. J.; He, Y. W.

In: Journal of Interferon and Cytokine Research, Vol. 15, No. 5, 01.01.1995, p. 447-454.

Research output: Contribution to journalArticle

Malek, T. R. ; Furse, R. K. ; Fleming, M. L. ; Fadell, A. J. ; He, Y. W. / Biochemical identity and characterization of the mouse interleukin-2 receptor β and γ(c) subunits. In: Journal of Interferon and Cytokine Research. 1995 ; Vol. 15, No. 5. pp. 447-454.
@article{d79530e419d9448985c6ccce0d00f9fc,
title = "Biochemical identity and characterization of the mouse interleukin-2 receptor β and γ(c) subunits",
abstract = "Although the mouse IL-2 receptor (IL-2R) β and γ(c) subunits have been identified by molecular cloning, the biochemical identity of these subunits has not yet been established. In the present study, the mouse IL-2R was biochemically characterized from cell lines expressing normal and aberrant IL-2R. Using novel monoclonal antibodies specific for the β or γ(c) subunits, we established that the M(r) of the β chain is 90,000-100,000 and that of the γ(c) subunit is 75,000-80,000. Analysis of transfected EL4 cells that expressed α, γ(c), and truncated β subunits or mutant EL4 cells, which selectively lacked cell surface γ(c), revealed that no other material migrated to a position on SDS-PAGE characteristic of IL-2/IL-2Rβ and IL- 2/IL-2Rγ(c) cross-linked complexes, respectively. Thus, the β and γ(c) subunits appear to be the sole IL-2R constituents of these IL-2 cross-linked complexes. The IL-2/IL-2Rγ(c), but not the IL-2/IL-2Rβ, complex exhibited enhanced mobility after SDS-polyacrylamide gel electrophoresis under nonreducing conditions, suggesting a more compact structure for γ(c) as a result of intrachain disulfide bonds. The primary posttranslational modification of the mouse β and γ(c) subunits is N-linked glycosylation. These biochemical studies reconcile past uncertainties concerning the subunit composition of the mouse IL-2R and are consistent with a model of the IL-2R containing only three subunits.",
author = "Malek, {T. R.} and Furse, {R. K.} and Fleming, {M. L.} and Fadell, {A. J.} and He, {Y. W.}",
year = "1995",
month = "1",
day = "1",
language = "English",
volume = "15",
pages = "447--454",
journal = "Journal of Interferon and Cytokine Research",
issn = "1079-9907",
publisher = "Mary Ann Liebert Inc.",
number = "5",

}

TY - JOUR

T1 - Biochemical identity and characterization of the mouse interleukin-2 receptor β and γ(c) subunits

AU - Malek, T. R.

AU - Furse, R. K.

AU - Fleming, M. L.

AU - Fadell, A. J.

AU - He, Y. W.

PY - 1995/1/1

Y1 - 1995/1/1

N2 - Although the mouse IL-2 receptor (IL-2R) β and γ(c) subunits have been identified by molecular cloning, the biochemical identity of these subunits has not yet been established. In the present study, the mouse IL-2R was biochemically characterized from cell lines expressing normal and aberrant IL-2R. Using novel monoclonal antibodies specific for the β or γ(c) subunits, we established that the M(r) of the β chain is 90,000-100,000 and that of the γ(c) subunit is 75,000-80,000. Analysis of transfected EL4 cells that expressed α, γ(c), and truncated β subunits or mutant EL4 cells, which selectively lacked cell surface γ(c), revealed that no other material migrated to a position on SDS-PAGE characteristic of IL-2/IL-2Rβ and IL- 2/IL-2Rγ(c) cross-linked complexes, respectively. Thus, the β and γ(c) subunits appear to be the sole IL-2R constituents of these IL-2 cross-linked complexes. The IL-2/IL-2Rγ(c), but not the IL-2/IL-2Rβ, complex exhibited enhanced mobility after SDS-polyacrylamide gel electrophoresis under nonreducing conditions, suggesting a more compact structure for γ(c) as a result of intrachain disulfide bonds. The primary posttranslational modification of the mouse β and γ(c) subunits is N-linked glycosylation. These biochemical studies reconcile past uncertainties concerning the subunit composition of the mouse IL-2R and are consistent with a model of the IL-2R containing only three subunits.

AB - Although the mouse IL-2 receptor (IL-2R) β and γ(c) subunits have been identified by molecular cloning, the biochemical identity of these subunits has not yet been established. In the present study, the mouse IL-2R was biochemically characterized from cell lines expressing normal and aberrant IL-2R. Using novel monoclonal antibodies specific for the β or γ(c) subunits, we established that the M(r) of the β chain is 90,000-100,000 and that of the γ(c) subunit is 75,000-80,000. Analysis of transfected EL4 cells that expressed α, γ(c), and truncated β subunits or mutant EL4 cells, which selectively lacked cell surface γ(c), revealed that no other material migrated to a position on SDS-PAGE characteristic of IL-2/IL-2Rβ and IL- 2/IL-2Rγ(c) cross-linked complexes, respectively. Thus, the β and γ(c) subunits appear to be the sole IL-2R constituents of these IL-2 cross-linked complexes. The IL-2/IL-2Rγ(c), but not the IL-2/IL-2Rβ, complex exhibited enhanced mobility after SDS-polyacrylamide gel electrophoresis under nonreducing conditions, suggesting a more compact structure for γ(c) as a result of intrachain disulfide bonds. The primary posttranslational modification of the mouse β and γ(c) subunits is N-linked glycosylation. These biochemical studies reconcile past uncertainties concerning the subunit composition of the mouse IL-2R and are consistent with a model of the IL-2R containing only three subunits.

UR - http://www.scopus.com/inward/record.url?scp=0029005081&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029005081&partnerID=8YFLogxK

M3 - Article

C2 - 7648447

VL - 15

SP - 447

EP - 454

JO - Journal of Interferon and Cytokine Research

JF - Journal of Interferon and Cytokine Research

SN - 1079-9907

IS - 5

ER -