Heterogeneity of protein phosphorylation in metastatic variants of B16 melanoma

R. G. Greig, L. Caltabiano, R. Reid, J. Feild, George Poste

Research output: Contribution to journalArticle

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Abstract

The polypeptide and phosphoprotein profiles of a spectrum of B16 melanoma clones of defined metastatic potential have been analyzed by two-dimensional gel electrophoresis. To accommodate the documented instability of metastatic properties in B16 clones, in vitro biochemical assays were always accompanied by in vivo assays of the metastatic behavior using replicate samples of the same clonal populations harvested on the same day. To exclude differences in polypeptide and phosphoprotein profiles resulting from inherent variation in electrophoretic measurements made at different times, polypeptides and phosphoproteins were analyzed in unison for every clone, and a series of clones was examined in parallel in each experiment. Also, samples were electrophoresed simultaneously using a custom-designed apparatus capable of accommodating 20 two-dimensional samples. When tested under these stringent conditions, the polypeptide profiles of B16 clones were indistinguishable. Significant qualitative and quantitative differences in phosphoprotein expression were detected in each clone, but no correlations were found between alterations in protein phosphorylation and metastatic potential. Over 200 discrete phosphoproteins were detected in each clone, but interclonal variation was confined to approximately 10 to 15 phosphoproteins. Expression of three phosphoproteins with the following molecular weights (in kilodaltons) and isoelectric points was strictly qualitative: pp96 (7.9); pp30 (8.2); and pp30 (8.8). In any given clone, they were present individually at equal intensities or were completely absent, but their expression was not coordinate. The data indicate that expression of polypeptide gene products is similar in B16 melanoma clones with widely differing metastatic abilities, but considerable clonal variability exists in posttranslational covalent modification of cell proteins. The possible contribution of protein phosphorylation and other posttranslational pathways in generating the extensive phenotypic heterogeneity observed in tumor cell subpopulations within the same tumor and in the rapid generation of new clonal variants with altered metastatic properties is discussed.

Original languageEnglish (US)
Pages (from-to)6057-6065
Number of pages9
JournalCancer Research
Volume43
Issue number12 I
StatePublished - 1983
Externally publishedYes

Fingerprint

Experimental Melanomas
Phosphoproteins
Clone Cells
Phosphorylation
Proteins
Peptides
Electrophoresis, Gel, Two-Dimensional
Isoelectric Point
Post Translational Protein Processing
Neoplasms
Molecular Weight
Gene Expression

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Greig, R. G., Caltabiano, L., Reid, R., Feild, J., & Poste, G. (1983). Heterogeneity of protein phosphorylation in metastatic variants of B16 melanoma. Cancer Research, 43(12 I), 6057-6065.

Heterogeneity of protein phosphorylation in metastatic variants of B16 melanoma. / Greig, R. G.; Caltabiano, L.; Reid, R.; Feild, J.; Poste, George.

In: Cancer Research, Vol. 43, No. 12 I, 1983, p. 6057-6065.

Research output: Contribution to journalArticle

Greig, RG, Caltabiano, L, Reid, R, Feild, J & Poste, G 1983, 'Heterogeneity of protein phosphorylation in metastatic variants of B16 melanoma', Cancer Research, vol. 43, no. 12 I, pp. 6057-6065.
Greig RG, Caltabiano L, Reid R, Feild J, Poste G. Heterogeneity of protein phosphorylation in metastatic variants of B16 melanoma. Cancer Research. 1983;43(12 I):6057-6065.
Greig, R. G. ; Caltabiano, L. ; Reid, R. ; Feild, J. ; Poste, George. / Heterogeneity of protein phosphorylation in metastatic variants of B16 melanoma. In: Cancer Research. 1983 ; Vol. 43, No. 12 I. pp. 6057-6065.
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