Characterization and subcellular localization of human Pmel 17/silver, a 110-kDa (pre)melanosomal membrane protein associated with 5,6,-dihydroxyindole-2-carboxylic acid (DHICA) converting activity.

Pmel 17 is preferentially expressed in pigment cells in a manner suggestive of involvement in melanin biosynthesis. The gene is identical to the silver (si) pigmentation locus in mice. We now produced a recombinant glutathione-S-transferase-human Pmel 17 infusion protein and raised polyclonal antibodies against it to confirm the ultrastructural location and presumed site of action predicted by the deduced primary structure of Pmel 17/silver, and to authenticate the specificity of the DHICA converting function as inherent to the silver-locus protein. Full-length Pmel 17 cDNA also produced in insect cells in a baculovirus expression vector to ensure that activity did not originate from a co-precipitated protein. Natural hPmel 17 from human melanoma cells has an approximate molecular size of 100 kDa. By immunoperoxidase electron microscopic cytochemistry, the antigen was localized to the limiting membranes of premelanosomes and presumed premelanogenic cytosolic vesicles and, to a minor extent, in the premelanosomal matrix. In an in vitro assay, both the natural and the recombinant Pmel 17 accelerated the conversion of DHICA to melanin. This activity was inhibited by the anti-Pmel 17 polyclonal antibodies, indicating that the acceleration of DHICA conversion by natural protein is genuine and cannot be due to contaminating complexed proteins. We suggest that in situ Pmel 17/silver is a component of a postulated premelanosomal/melanosomal complex of membrane-bound melanogenic oxidoreductive enzymes and cofactors, in analogy to the electron transfer chain in mitochondria.

[1]  A. Houghton,et al.  Intracellular sorting and targeting of melanosomal membrane proteins: identification of signals for sorting of the human brown locus protein, gp75 , 1995, The Journal of cell biology.

[2]  Dorothy Bennett,et al.  Mouse silver mutation is caused by a single base insertion in the putative cytoplasmic domain of Pmel 17 , 1995, Nucleic Acids Res..

[3]  B. Kwon,et al.  Characterization of mouse Pmel 17 gene and silver locus. , 1994, Pigment cell research.

[4]  G. Imokawa,et al.  Tyrosinase related protein 1 (TRP1) functions as a DHICA oxidase in melanin biosynthesis. , 1994, The EMBO journal.

[5]  Richard P. Woychik,et al.  Agouti protein is an antagonist of the melanocyte-stimulating-hormone receptor , 1994, Nature.

[6]  E. Rosengren,et al.  The mouse brown (b) locus protein functions as a dopachrome tautomerase. , 1994, Pigment cell research.

[7]  C. Figdor,et al.  Molecular characterization of the melanocyte lineage-specific antigen gp100. , 1994, The Journal of biological chemistry.

[8]  S. Orlow,et al.  High-molecular-weight forms of tyrosinase and the tyrosinase-related proteins: evidence for a melanogenic complex. , 1994, The Journal of investigative dermatology.

[9]  K. Urabe,et al.  Functional analysis of the slaty gene product (TRP2) as dopachrome tautomerase and the effect of a point mutation on its catalytic function. , 1994, Biochemical and biophysical research communications.

[10]  S. Orlow,et al.  Identification of a melanosomal matrix protein encoded by the murine si (silver) locus using "organelle scanning". , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[11]  K. Urabe,et al.  A new enzymatic function in the melanogenic pathway. The 5,6-dihydroxyindole-2-carboxylic acid oxidase activity of tyrosinase-related protein-1 (TRP1). , 1994, The Journal of biological chemistry.

[12]  K. Sakaguchi,et al.  Identification of a human melanoma antigen recognized by tumor-infiltrating lymphocytes associated with in vivo tumor rejection. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[13]  R. Henderson,et al.  Identification of a peptide recognized by five melanoma-specific human cytotoxic T cell lines. , 1994, Science.

[14]  C. Figdor,et al.  Melanocyte lineage-specific antigens recognized by monoclonal antibodies NKI-beteb, HMB-50, and HMB-45 are encoded by a single cDNA. , 1993, The American journal of pathology.

[15]  E. Rosengren,et al.  The mouse brown (b) locus protein has dopachrome tautomerase activity and is located in lysosomes in transfected fibroblasts. , 1993, Journal of cell science.

[16]  S. Orlow,et al.  Identification of a mammalian melanosomal matrix glycoprotein. , 1993, The Journal of investigative dermatology.

[17]  D. Taatjes,et al.  HMB-45 antibody demonstrates melanosome specificity by immunoelectron microscopy. , 1993, Archives of pathology & laboratory medicine.

[18]  B. Kwon,et al.  Pigmentation genes: the tyrosinase gene family and the pmel 17 gene family. , 1993, The Journal of investigative dermatology.

[19]  S. Orlow,et al.  Subcellular distribution of tyrosinase and tyrosinase-related protein-1: implications for melanosomal biogenesis. , 1993, The Journal of investigative dermatology.

[20]  G. Wistow,et al.  The cDNA RPE1 and monoclonal antibody HMB-50 define gene products preferentially expressed in retinal pigment epithelium. , 1992, Experimental eye research.

[21]  I. Jackson,et al.  Light is a dominant mouse mutation resulting in premature cell death , 1992, Nature Genetics.

[22]  E. Kaiserling,et al.  Ultrastructural localization of HMB‐45 binding sites , 1991, Journal of cutaneous pathology.

[23]  D. Barton,et al.  A melanocyte-specific gene, Pmel 17, maps near the silver coat color locus on mouse chromosome 10 and is in a syntenic region on human chromosome 12. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. Shibahara,et al.  The monoclonal antibodies TMH-1 and TMH-2 specifically bind to a protein encoded at the murine b-locus, not to the authentic tyrosinase encoded at the c-locus. , 1991, The Journal of investigative dermatology.

[25]  K. Agata,et al.  Complete sequence and expression of a cDNA encoding a chicken 115-kDa melanosomal matrix protein. , 1991, Pigment cell research.

[26]  R. Halaban,et al.  Murine and human b locus pigmentation genes encode a glycoprotein (gp75) with catalase activity. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[27]  R. Halaban,et al.  Basic fibroblast growth factor from human keratinocytes is a natural mitogen for melanocytes , 1988, The Journal of cell biology.

[28]  I. Jackson,et al.  A cDNA encoding tyrosinase-related protein maps to the brown locus in mouse. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[29]  D. Ruiter,et al.  A monoclonal antibody specific for cells of the melanocyte lineage. , 1988, The American journal of pathology.

[30]  David W. Miller,et al.  An Insect Baculovirus Host-Vector System For High-Level Expression of Foreign Genes , 1986 .

[31]  S. Pomerantz,et al.  Tyrosinase activity and abundance in Cloudman melanoma cells. , 1984, Archives of biochemistry and biophysics.

[32]  T. Fitzpatrick,et al.  Characterization of a new melanosomal structural component--the vesiculoglobular body--by conventional transmission, high-voltage, and scanning electron microscopy. , 1974, Journal of ultrastructure research.

[33]  P. Novikoff,et al.  STUDIES ON MICROPEROXISOMES V. ARE MICROPEROXISOMES UBIQUITOUS IN MAMMALIAN CELLS? , 1973, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[34]  W. C. Quevedo,et al.  An analysis of the light mutation of coat color in mice , 1958 .