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Presence of a laminin-binding chondroitin sulfate proteoglycan at the cell surface of a human melanoma cell Mel-85

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Abstract

Working with Mel-85 (a human melanoma cell line), we have been able to detect a laminin-binding molecule with an apparent molecular mass of 100/110 kDa (Mel-85-LBM). Reduction with β-mercaptoethanol decreases its molecular mass but does not affect its ability to bind laminin. This laminin interaction seems to be very specific since Mel-85-LBM binds laminin, but not fibronectin, vitronectin or type I collagen in affinity chromatography experiments. The molecule has a negative net charge at physiological pH and binds laminin in a divalent cation dependent way. Mel-85-LBM was metabolically radiolabeled with sodium [35S]-sulfate and chemical β-elimination of purified Mel-85-LBM releases chondroitin sulfate chains. Mel-85-LBM is also sensitive to chondroitinase ABC digestion. These findings show that this molecule is a chondroitin sulfate proteoglycan. The location of this proteoglycan at the cell surface is evidenced by experiments using a polyclonal antiserum raised against purified Mel-85LBM, that specifically reacts with just one molecule by western blotting among Mel-85 total cell extract as well as produces a positive signal by flow cytometry and a fluorescence profile of Mel-85 cells adhered on laminin.

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Authors and Affiliations

  1. Ludwig Institute for Cancer Research, São Paulo, Brazil

    Maria Carolina Q.B. Elias, Silvio S. Veiga, Waldemiro Gremski & Ricardo R. Brentani

  2. Department of Cell Biology, Universidade Federal do Paraná, Curitiba, PR, Brazil

    Silvio S. Veiga & Waldemiro Gremski

  3. Department of Biochemistry, Escola Paulista de Medicina, UNIFESP, São Paulo, SP, Brazil

    Marimélia A. Porcionatto & Helena B. Nader

Authors
  1. Maria Carolina Q.B. Elias
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  2. Silvio S. Veiga
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  3. Waldemiro Gremski
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  4. Marimélia A. Porcionatto
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  5. Helena B. Nader
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  6. Ricardo R. Brentani
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Elias, M.C.Q., Veiga, S.S., Gremski, W. et al. Presence of a laminin-binding chondroitin sulfate proteoglycan at the cell surface of a human melanoma cell Mel-85. Mol Cell Biochem 197, 39–48 (1999). https://doi.org/10.1023/A:1006952731037

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  • DOI: https://doi.org/10.1023/A:1006952731037

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