PURIFICATION AND ANALYSIS OF GROWTH-REGULATING PROTEINS SECRETED BY A HUMAN-MELANOMA CELL-LINE

被引：49

作者：

APFEL, R ^{[1
]}

LOTTSPEICH, F ^{[1
]}

HOPPE, J ^{[1
]}

BEHL, C ^{[1
]}

DURR, G ^{[1
]}

BOGDAHN, U ^{[1
]}

机构：

[1] DEPT NEUROL,TUMORBIOL LAB,JOSEF SCHNEIDER STR 11,W-8700 WURZBURG,GERMANY

来源：

MELANOMA RESEARCH | 1992年 / 2卷 / 5-6期

关键词：

ENDOZEPINE; MALIGNANT MELANOMA; MELANOMA INHIBITING ACTIVITY; BETA-2-MICROGLOBULIN; TIMP-2; UBIQUITIN;

D O I：

10.1097/00008390-199212000-00006

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

Supernatants of a human malignant cell line established from a CNS metastasis, contained several proteins with putative growth regulating functions. BioGel P-10 gel filtration chromatography, reverse phase HPLC purification, and amino-terminal sequencing of purified peptides resulted in characterization of beta2-microglobulin (beta2M, 10 kD), ubiquitin (6 kD), and tissue inhibitor of metalloproteinases 2 (TIMP-2, 21 kD). In addition, CNBr cleavage and purification of resulting peptides revealed diazepam binding inhibitor (DBI, 8 kD) and melanoma inhibiting activity (MIA, 11 kD). The secretion of beta2M as part of the HLA-class I complex may be related to impaired autologous anti-tumour immune function; ubiquitin may play a role in activation or deactivation of extracellular proteins or cell-cell interactions. As HTZ-19 cells respond in a dose-dependent manner to midozolam, DBI may interfere with growth regulation mediated by diazepam receptor sites. In a collagenolytic assay, TIMP-2 interfered with metalloproteinase functions, which are required for degradation of collagen type IV and organotopic metastasis. MIA is clearly associated with a proliferation inhibiting effect on HTZ-19 cells. In conclusion, although this tumour shows a degree of progression, several proteins with putative functions at different cellular levels were identified, related to proliferation as well as to the type of metastasis.

引用

页码：327 / 336

页数：10

共 47 条

[31]

Pawlikowski M., Kunert-Radel J., Radel A., Stepien H., Inhibition of cell proliferation of human gliomas by benzodiazepines in vitro, Acta Neurol Scand, 77, pp. 231-233, (1988)

[32]

Cantor E.H., Kenessey A., Gemenul G., Spector S., Interaction of calcium channel blockers with non-neuronal benzodiazepine binding sites, Proc Natl Acad Sci., 81, pp. 1549-1552, (1984)

[33]

Sjuholm A., Funakoshi A., Efendic S., Ostenson C.G., Hellerstrum C., Long term inhibitory effects of pancreastatin and diazepam binding inhibitor on pancreatic b-cell deoxyribonucleic acid replication, polyamine content and insulin secretion, Endocrinology, 128, pp. 3277-3282, (1991)

[34]

Ciechanover A., Heller H., Katz-Etzion R., Herskho A., Activation of the heat-stable polypeptide of the ATP-dependent proteolytic system, Proc Natl Acad Sci., 78, pp. 761-765, (1981)

[35]

Ciechanover A., Heller H., Elias S., Haas A.L., Herskho A., Atp-dependent conjugation of reticulocyte proteins with the polypeptide required for protein degradation, Proc Natl Acad Sci., 77, pp. 1365-1368, (1980)

[36]

Nickel B.E., Davie J.R., Structure of polyubiquitinated histone H2A, Biochemistry, 28, pp. 964-968, (1989)

[37]

Nickel B.E., Allis C.D., David J.R., Ubiquitinated histone H2B is preferentially located in transcriptionally active chromatin, Biochemistry, 28, pp. 958-963, (1989)

[38]

Jentsch S., McGrath J.P., Varshavsky A., The yeast DNA repair gene rad6 encodes a ubiquitin-conjugating enzyme, Nature, 329, pp. 131-134, (1987)

[39]

Goeble M.G., Yochem J., Jentsch S., McGrath J., Vashavsky A., Byers B., The yeast cell cycle gene CDC 34 encodes a ubiquitin-conjugating enzyme, Science, 241, pp. 1331-1335, (1988)

[40]

Jentsch S., Seufert W., Hauser H.P., Genetic analysis of the ubiquitin system, Biochim Biophys Acta., 1089, pp. 127-139, (1991)

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