Premium
Improved detection of labile cell‐surface components with zinc chloride‐aprotinin: Demonstration of glycoprotein differences in K‐1735 metastatic melanoma variants
Author(s) -
Rieber Manuel,
Rieber Mary S.
Publication year - 1985
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910360517
Subject(s) - glycoprotein , melanoma , labelling , zinc , cell , molecular mass , chemistry , aprotinin , biochemistry , p glycoprotein , microbiology and biotechnology , biology , cancer research , medicine , enzyme , multiple drug resistance , antibiotics , organic chemistry
Metabolic labelling of K‐1735 melanoma variants with 3 H‐glucosamine and cell harvesting with the commonly used protease inhibitor phenylmethylsulfonylfluoride revealed a Triton‐insoluble fibronection‐like 230 kd component in poorly metastatic cells. This component was not evident in highly metastatic cells. Significantly improved surface labelling and detection of the 230 kd glycoprotein in the highly metastatic variant was achieved by zinc chloride‐aprotinin treatment of cells prior to harvesting. This procedure also revealed an increase in a trypsin‐sensitive glycoprotein of higher molecular weight in the Triton‐insoluble fraction of the highly metastatic cell variant. Glycoprotein labelling in this fraction showed an electrophoretic pattern strongly resembling that reported by others for the high‐molecular‐weight human melanoma‐associated glycoprotein complex. The differential detection of the high‐molecular‐weight glycoprotein species in melanoma variants with differing metastatic abilities in an animal model provides a means of studying their possible relevance to metastatic melanoma. Our data also suggest that zinc chlorideaprotinin can be used to improve the detection of labile cell‐surface components.