The beta isoform of protein kinase C stimulates human melanogenesis by activating tyrosinase in pigment cells.
We have investigated the role of protein kinase C (PKC) in human melanogenesis. The level of PKC activity paralleled the total melanin content in cultured newborn melanocytes. Activation of PKC by treatment with 5 x 10(-7) M phorbol dibutyrate acutely caused a doubling in the activity of tyrosinase, the rate-limiting enzyme in melanogenesis, known to correlate directly with melanin synthesis in these cells. When PKC was depleted to 5-10% of initial levels, there was a 40-50% parallel reduction in tyrosinase activity; and regeneration of PKC activity was associated with the recovery of tyrosinase activity. By Northern blot analysis, the alpha and beta but not the gamma isoforms were detectable in melanocytes. By Western blot analysis, the racially determined pigment level in cultured melanocytes correlated with PKC-beta protein expression. In a pigmented human melanoma line (P-MM4, 20-30 ng melanin/micrograms protein)and its nonpigmented subclone (NP-MM4, undetectable melanin), PKC-alpha mRNA was expressed in both, whereas PKC-beta mRNA was detectable only in P-MM4 cells. Tyrosinase protein level was comparable in both cell lines. When NP-MM4 cell lysate was incubated with melanocyte lysate known to contain PKC-beta, tyrosinase activity per microgram of melanocyte protein in the combined lysate increased, consistent with activation of the previously inactive tyrosinase of NP-MM4 origin. Moreover, NP-MM4 cells transiently transfected with PKC-beta cDNA increased tyrosinase activity from undetectable to detectable levels. These combined data show that PKC-beta regulates human melanogenesis by activating tyrosinase.
Park
H Y
HY
Department of Dermatology, Boston University School of Medicine, Massachusetts 02118.
Russakovsky
V
V
Ohno
S
S
Gilchrest
B A
BA
D015152:Blotting, Northern; D002455:Cell Division; D002478:Cells, Cultured; D015342:DNA Probes; D004789:Enzyme Activation; D006801:Humans; D007231:Infant, Newborn; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008543:Melanins / Q000096:biosynthesis / Q000378:metabolism*; D008544:Melanocytes / Q000166:cytology / Q000201:enzymology*; D014442:Monophenol Monooxygenase / Q000235:genetics / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012867:Skin / Q000201:enzymology*
D015342:DNA Probes; D007527:Isoenzymes; D008543:Melanins; D012313:RNA; D014442:Monophenol Monooxygenase; D011493:Protein Kinase C