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Subcellular fractionation of cultured normal human melanocytes: New insights into the relationship of melanosomes with lysosomes and peroxisomes |
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| Authors: | Nico P.M. Smit Carlo W.T. van Roermund Hans M.F.G. Aerts Judith C. Heikoop Marlene Van den Berg Stan Pavel Ronald J.A. Wanders |
| Affiliation: | 1. Department of Dermatology, University of Amsterdam, Academic Medical Center, Amsterdam, Netherlands;2. Department of Paediatrics, University of Amsterdam, Academic Medical Center, Amsterdam, Netherlands;3. E.C. Slater Institute for Biochemical Research, University of Amsterdam, Academic Medical Center, Amsterdam, Netherlands;4. Department of Dermatology, University Hospital, Leiden, Netherlands |
| Abstract: | In order to obtain information on the disputed nature of melanosomes a comparison was made between the localization of melanosomal markers with those of other well-defined subcellular organelles such as lysosomes and peroxisomes. The distribution of marker enzymes was studied using two different density gradient systems. i.e., Percoll and Nycodenz. Furthermore, the subcellular localization of various types of antigens was analyzed using indirect immunofluorescence and immuno-electron microscopy. All methods revealed the existence of partial co-localization of melanosomal and lysosomal proteins and different localization of peroxisomal markers. The results suggest that melanosomes may share a common origin with lysosomal structures. |
| Keywords: | Melanocyte Melanosome Lysosome Peroxisome DHAPAT acyl CoA dihydroxyacetonephosphate acyltransferase GDH glutamate dehydrogenase GERL Golgi-endoplasmic reticulum-lysosome complex IBMX isobutylmethylxanthine LDH lactate dehydrogenase Mc melanocytes MoAbs monoclonal antibodies MOPS morpholinopropane sulphonic acid PMP peroxisomal membrane protein PNS post-nuclear supernatant TPA 12-O-tetradecanoyl phorbol 13-acetate |
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