Ligand-induced Phosphorylation of a Murine Tumor Surface Protein (TSP-180) Associated with Metastatic Phenotype

A. Sacchi, R. Falcioni, G. Piaggio, M. A. Gianfelice, N. Perrotti, Stephen Kennel

Research output: Contribution to journalArticle

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Abstract

A tumor surface protein (TSP-180) has been identified on murine lung carcinomas using two monoclonal antibodies (MoAbs) (135-13C and 346-11 A). Quantitative analysis of TSP-180 on 3LL variants maintained either in vitro or in vivo indicates that TSP-180 is highly expressed in highly malignant metastatic cells. In reducing conditions, sodium dodecyl sulfate-polyacrylamide gel electrophoresis banding patterns of TSP-180 obtained with MoAb 135-13C from cell lysates of 3LL metastatic cells show three proteins migrating to A#r 204,000,134,000, and 116,000. In the same experimental conditions MoAb 135-13C precipitates from low metastasizing ones only one band, corresponding to the lower molecular weight (Mr 116,000). All bands of TSP-180 observed in 3LL variants are labeled by lactoperoxidase-catalyzed radioiodination of viable cells, incorporate “PO and contain carbohydrates, as judged by binding to wheat germ agglutinin. These results indicate that all proteins have external exposure on the cell surface and that at least some of TSP-180 proteins could be differentially regulated in different tumor cells (highly metastatic versus low metastatic). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis banding patterns and immunoblots obtained from cell lysates of 3LL variants by using a monoclonal antibody to phosphotyro-sine (IG-2) indicate that this MoAb recognizes proteins migrating with molecular weights identical to those reported for TSP-180. Moreover, the immunoblots of solubilized immunocomplex, obtained from cell lysates of 3LL variants by using MoAb 135-13C, demonstrate that MoAb IG-2 specifically reacts with TSP-180 proteins. Experiments undertaken in order to assess if some or all of TSP-180 proteins have tyrosine kinase activity demonstrate that MoAb 135-13C binding to the cell surface induces specific phosphorylation of the M, 204,000 protein of TSP-180. Phosphoaminoacid analysis of the ligand-induced phosphorylated protein (pp204) demonstrates that this protein is phosphorylated at serine and tyrosine. Results reported lead us to hypothesize that TSP-180 is involved in growth-regulation mechanisms and that its high expression on cells with more malignant phenotype could be responsible for a proliferative advantage of such tumor clones.

Original languageEnglish (US)
Pages (from-to)2615-2620
Number of pages6
JournalCancer Research
Volume49
Issue number10
StatePublished - May 15 1989
Externally publishedYes

Fingerprint

Integrin alpha6beta4
Membrane Proteins
Phosphorylation
Ligands
Phenotype
Neoplasms
Proteins
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Molecular Weight
Monoclonal Antibodies
Lactoperoxidase
Wheat Germ Agglutinins

All Science Journal Classification (ASJC) codes

  • Cancer Research
  • Oncology

Cite this

Sacchi, A., Falcioni, R., Piaggio, G., Gianfelice, M. A., Perrotti, N., & Kennel, S. (1989). Ligand-induced Phosphorylation of a Murine Tumor Surface Protein (TSP-180) Associated with Metastatic Phenotype. Cancer Research, 49(10), 2615-2620.

Ligand-induced Phosphorylation of a Murine Tumor Surface Protein (TSP-180) Associated with Metastatic Phenotype. / Sacchi, A.; Falcioni, R.; Piaggio, G.; Gianfelice, M. A.; Perrotti, N.; Kennel, Stephen.

In: Cancer Research, Vol. 49, No. 10, 15.05.1989, p. 2615-2620.

Research output: Contribution to journalArticle

Sacchi, A, Falcioni, R, Piaggio, G, Gianfelice, MA, Perrotti, N & Kennel, S 1989, 'Ligand-induced Phosphorylation of a Murine Tumor Surface Protein (TSP-180) Associated with Metastatic Phenotype', Cancer Research, vol. 49, no. 10, pp. 2615-2620.
Sacchi A, Falcioni R, Piaggio G, Gianfelice MA, Perrotti N, Kennel S. Ligand-induced Phosphorylation of a Murine Tumor Surface Protein (TSP-180) Associated with Metastatic Phenotype. Cancer Research. 1989 May 15;49(10):2615-2620.
Sacchi, A. ; Falcioni, R. ; Piaggio, G. ; Gianfelice, M. A. ; Perrotti, N. ; Kennel, Stephen. / Ligand-induced Phosphorylation of a Murine Tumor Surface Protein (TSP-180) Associated with Metastatic Phenotype. In: Cancer Research. 1989 ; Vol. 49, No. 10. pp. 2615-2620.
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abstract = "A tumor surface protein (TSP-180) has been identified on murine lung carcinomas using two monoclonal antibodies (MoAbs) (135-13C and 346-11 A). Quantitative analysis of TSP-180 on 3LL variants maintained either in vitro or in vivo indicates that TSP-180 is highly expressed in highly malignant metastatic cells. In reducing conditions, sodium dodecyl sulfate-polyacrylamide gel electrophoresis banding patterns of TSP-180 obtained with MoAb 135-13C from cell lysates of 3LL metastatic cells show three proteins migrating to A#r 204,000,134,000, and 116,000. In the same experimental conditions MoAb 135-13C precipitates from low metastasizing ones only one band, corresponding to the lower molecular weight (Mr 116,000). All bands of TSP-180 observed in 3LL variants are labeled by lactoperoxidase-catalyzed radioiodination of viable cells, incorporate “PO and contain carbohydrates, as judged by binding to wheat germ agglutinin. These results indicate that all proteins have external exposure on the cell surface and that at least some of TSP-180 proteins could be differentially regulated in different tumor cells (highly metastatic versus low metastatic). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis banding patterns and immunoblots obtained from cell lysates of 3LL variants by using a monoclonal antibody to phosphotyro-sine (IG-2) indicate that this MoAb recognizes proteins migrating with molecular weights identical to those reported for TSP-180. Moreover, the immunoblots of solubilized immunocomplex, obtained from cell lysates of 3LL variants by using MoAb 135-13C, demonstrate that MoAb IG-2 specifically reacts with TSP-180 proteins. Experiments undertaken in order to assess if some or all of TSP-180 proteins have tyrosine kinase activity demonstrate that MoAb 135-13C binding to the cell surface induces specific phosphorylation of the M, 204,000 protein of TSP-180. Phosphoaminoacid analysis of the ligand-induced phosphorylated protein (pp204) demonstrates that this protein is phosphorylated at serine and tyrosine. Results reported lead us to hypothesize that TSP-180 is involved in growth-regulation mechanisms and that its high expression on cells with more malignant phenotype could be responsible for a proliferative advantage of such tumor clones.",
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N2 - A tumor surface protein (TSP-180) has been identified on murine lung carcinomas using two monoclonal antibodies (MoAbs) (135-13C and 346-11 A). Quantitative analysis of TSP-180 on 3LL variants maintained either in vitro or in vivo indicates that TSP-180 is highly expressed in highly malignant metastatic cells. In reducing conditions, sodium dodecyl sulfate-polyacrylamide gel electrophoresis banding patterns of TSP-180 obtained with MoAb 135-13C from cell lysates of 3LL metastatic cells show three proteins migrating to A#r 204,000,134,000, and 116,000. In the same experimental conditions MoAb 135-13C precipitates from low metastasizing ones only one band, corresponding to the lower molecular weight (Mr 116,000). All bands of TSP-180 observed in 3LL variants are labeled by lactoperoxidase-catalyzed radioiodination of viable cells, incorporate “PO and contain carbohydrates, as judged by binding to wheat germ agglutinin. These results indicate that all proteins have external exposure on the cell surface and that at least some of TSP-180 proteins could be differentially regulated in different tumor cells (highly metastatic versus low metastatic). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis banding patterns and immunoblots obtained from cell lysates of 3LL variants by using a monoclonal antibody to phosphotyro-sine (IG-2) indicate that this MoAb recognizes proteins migrating with molecular weights identical to those reported for TSP-180. Moreover, the immunoblots of solubilized immunocomplex, obtained from cell lysates of 3LL variants by using MoAb 135-13C, demonstrate that MoAb IG-2 specifically reacts with TSP-180 proteins. Experiments undertaken in order to assess if some or all of TSP-180 proteins have tyrosine kinase activity demonstrate that MoAb 135-13C binding to the cell surface induces specific phosphorylation of the M, 204,000 protein of TSP-180. Phosphoaminoacid analysis of the ligand-induced phosphorylated protein (pp204) demonstrates that this protein is phosphorylated at serine and tyrosine. Results reported lead us to hypothesize that TSP-180 is involved in growth-regulation mechanisms and that its high expression on cells with more malignant phenotype could be responsible for a proliferative advantage of such tumor clones.

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