Reconstitution of the multiprotein complex of pp60(src), hsp90, and p50 in a cell-free system

K. A. Hutchison, B. K. Brott, J. H. De Leon, G. H. Perdew, R. Jove, W. B. Pratt

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

A rabbit reticulocyte lysate system that has been used to reconstitute functional complexes between steroid receptors and the 90-kDa heat shock protein (hsp90) has been used here to form complexes between the pp60(src) tyrosine kinase and hsp90. Reticulocyte lysate forms complexes between hsp90 and a temperature-sensitive mutant of Rous sarcoma virus pp60(v-src), which is normally present in cytosol virtually entirely in the multiprotein complex form. In addition, hsp90 in the lysate complexes with wild-type pp60(v-src), of which only a small portion is normally recovered in cytosol in the native multiprotein complex, and with the cellular homolog, pp60(c-src), which has never been recovered in cytosol in the form of a native multiprotein complex with hsp90. Moreover, the reticulocyte lysate-reconstituted complex also contains the 50-kDa phosphoprotein component of the native pp60(v-src) multiprotein complex. The native and reconstituted pp60(src)-hsp90 complexes have similar thermal stability and, like steroid receptor heterocomplexes, they are stabilized by molybdate. As previously shown with reticulocyte lysate-reconstituted steroid receptor heteroprotein complexes, the reconstituted pp60(src) multiprotein complex contains hsp70, which is a major candidate for providing the protein unfoldase activity required for hsp90 association.

Original languageEnglish (US)
Pages (from-to)2902-2908
Number of pages7
JournalJournal of Biological Chemistry
Volume267
Issue number5
StatePublished - Jan 1 1992

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Multiprotein Complexes
Cell-Free System
Oncogene Protein pp60(v-src)
Reticulocytes
Steroid Receptors
Cytosol
Proto-Oncogene Proteins pp60(c-src)
HSP90 Heat-Shock Proteins
Rous sarcoma virus
src-Family Kinases
Phosphoproteins
Heat-Shock Proteins
Viruses
Thermodynamic stability
Hot Temperature
Rabbits
Temperature
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Hutchison, K. A., Brott, B. K., De Leon, J. H., Perdew, G. H., Jove, R., & Pratt, W. B. (1992). Reconstitution of the multiprotein complex of pp60(src), hsp90, and p50 in a cell-free system. Journal of Biological Chemistry, 267(5), 2902-2908.
Hutchison, K. A. ; Brott, B. K. ; De Leon, J. H. ; Perdew, G. H. ; Jove, R. ; Pratt, W. B. / Reconstitution of the multiprotein complex of pp60(src), hsp90, and p50 in a cell-free system. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 5. pp. 2902-2908.
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abstract = "A rabbit reticulocyte lysate system that has been used to reconstitute functional complexes between steroid receptors and the 90-kDa heat shock protein (hsp90) has been used here to form complexes between the pp60(src) tyrosine kinase and hsp90. Reticulocyte lysate forms complexes between hsp90 and a temperature-sensitive mutant of Rous sarcoma virus pp60(v-src), which is normally present in cytosol virtually entirely in the multiprotein complex form. In addition, hsp90 in the lysate complexes with wild-type pp60(v-src), of which only a small portion is normally recovered in cytosol in the native multiprotein complex, and with the cellular homolog, pp60(c-src), which has never been recovered in cytosol in the form of a native multiprotein complex with hsp90. Moreover, the reticulocyte lysate-reconstituted complex also contains the 50-kDa phosphoprotein component of the native pp60(v-src) multiprotein complex. The native and reconstituted pp60(src)-hsp90 complexes have similar thermal stability and, like steroid receptor heterocomplexes, they are stabilized by molybdate. As previously shown with reticulocyte lysate-reconstituted steroid receptor heteroprotein complexes, the reconstituted pp60(src) multiprotein complex contains hsp70, which is a major candidate for providing the protein unfoldase activity required for hsp90 association.",
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Hutchison, KA, Brott, BK, De Leon, JH, Perdew, GH, Jove, R & Pratt, WB 1992, 'Reconstitution of the multiprotein complex of pp60(src), hsp90, and p50 in a cell-free system', Journal of Biological Chemistry, vol. 267, no. 5, pp. 2902-2908.

Reconstitution of the multiprotein complex of pp60(src), hsp90, and p50 in a cell-free system. / Hutchison, K. A.; Brott, B. K.; De Leon, J. H.; Perdew, G. H.; Jove, R.; Pratt, W. B.

In: Journal of Biological Chemistry, Vol. 267, No. 5, 01.01.1992, p. 2902-2908.

Research output: Contribution to journalArticle

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T1 - Reconstitution of the multiprotein complex of pp60(src), hsp90, and p50 in a cell-free system

AU - Hutchison, K. A.

AU - Brott, B. K.

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AU - Jove, R.

AU - Pratt, W. B.

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N2 - A rabbit reticulocyte lysate system that has been used to reconstitute functional complexes between steroid receptors and the 90-kDa heat shock protein (hsp90) has been used here to form complexes between the pp60(src) tyrosine kinase and hsp90. Reticulocyte lysate forms complexes between hsp90 and a temperature-sensitive mutant of Rous sarcoma virus pp60(v-src), which is normally present in cytosol virtually entirely in the multiprotein complex form. In addition, hsp90 in the lysate complexes with wild-type pp60(v-src), of which only a small portion is normally recovered in cytosol in the native multiprotein complex, and with the cellular homolog, pp60(c-src), which has never been recovered in cytosol in the form of a native multiprotein complex with hsp90. Moreover, the reticulocyte lysate-reconstituted complex also contains the 50-kDa phosphoprotein component of the native pp60(v-src) multiprotein complex. The native and reconstituted pp60(src)-hsp90 complexes have similar thermal stability and, like steroid receptor heterocomplexes, they are stabilized by molybdate. As previously shown with reticulocyte lysate-reconstituted steroid receptor heteroprotein complexes, the reconstituted pp60(src) multiprotein complex contains hsp70, which is a major candidate for providing the protein unfoldase activity required for hsp90 association.

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