Vol. 273, Issue 3, 1605-1612, January 16, 1998
Purification and Characterization of an Allergy-induced
Melanogenic Stimulating Factor in Brownish Guinea Pig Skin
Genji
Imokawa
,
K.
Higuchi, and
Y.
Yada¶
From the Biological Science Laboratories and
¶ Tokyo Research Laboratories, Kao Corporation, Ichikaimachi 2606, Tochigi 321-34, and Bunka 2-chome, Sumida-ku, Tokyo 131 Japan
We have demonstrated recently that
phenylazonaphthol (PAN) allergy-induced hyperpigmentation in brownish
guinea pig skin is associated with the concomitant appearance of a
melanogenic soluble factor(s) that activates the intracellular signal
transduction system, including phosphatidylinositol turnover subsequent
to ligand-receptor binding in cultured guinea pig melanocytes. In this
study we have purified and characterized the PAN-induced melanogenic
stimulating factor (PIMSF) that occurs in allergy-associated hyperpigmented skin. By successive column chromatography on TSK 2000SW,
Mono Q, and octadecyl-NPR, the PIMSF was purified to homogeneity with a
single band of apparent molecular mass of 7.9 kDa by sodium dodecyl
sulfate-polyacrylamide gel electrophoresis. The specific bioactivity of
PIMSF increased by 5,195-fold over the original skin homogenate. In
cultured guinea pig melanocytes, this purified PIMSF had the potential
of activating an intracellular signal transduction system such as
inositol 1,4,5-trisphosphate formation and intracellular calcium levels
through a pertussis toxin-sensitive G protein-coupled receptor. PIMSF
consistently caused a rapid translocation of cytosolic protein kinase C
(PKC) to membrane-bound PKC within 5 min of treatment with a return to
the basal level after 120 min. The stimulating effects of PIMSF on
proliferation and melanization of cultured guinea pig melanocytes were
abolished completely by a PKC down-regulating agent (phorbol
12,13-dibutyrate). PIMSF was similar in molecular mass to rat
growth-related oncogene 
(GRO-; molecular mass of 7.9 kDa) on
sodium dodecyl sulfate-polyacrylamide gel electrophoresis and had
immunocross-reactivity with GRO- upon Western immune blotting
analysis. Further, the stimulatory effect of purified PIMSF on DNA
synthesis of cultured guinea pig melanocytes was suppressed markedly by
the addition of anti-rat GRO- antibody, implying that the PIMSF is
apparently identical to GRO-. These findings suggest that PAN
allergy provides a new mechanism of hyperpigmentation in which
biological factors such as the GRO- superfamily generated within
allergy-induced skin stimulate melanocytes through activation of the
PKC-related signal transduction pathway.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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