Human phenylalanine hydroxylase is activated by H2O2: a novel mechanism for increasing the L-tyrosine supply for melanogenesis in melanocytes.
Epidermal phenylalanine hydroxylase (PAH) produces L-tyrosine from the essential amino acid L-phenylalanine supporting melanogenesis in human melanocytes. Those PAH activities increase linearly in the different skin phototypes I-VI (Fitzpatrick classification) and also increase up to 24h after UVB light with only one minimal erythemal dose. Since UVB generates also H(2)O(2), we here asked the question whether this reactive oxygen species could influence the activity of pure recombinant human PAH. Under saturating conditions with the substrate L-phenylalanine (1x10(-3)M), the V(max) for enzyme activity increased 4-fold by H(2)O(2) (>2.0x10(-3)M). Lineweaver-Burk analysis identified a mixed activation mechanism involving both the regulatory and catalytic domains of PAH. Hyperchem molecular modelling and Deep View analysis support oxidation of the single Trp(120) residue to 5-OH-Trp(120) by H(2)O(2) causing a conformational change in the regulatory domain. PAH was still activated by H(2)O(2) in the presence of the electron donor/cofactor 6(R)-L-erythro-5,6,7,8-tetrahydrobiopterin despite slow oxidation of this cofactor. In vivo FT-Raman spectroscopy confirmed decreased epidermal phenylalanine in association with increased tyrosine after UVB exposure. Hence, generation of H(2)O(2) by UVB can activate epidermal PAH leading to an increased L-tyrosine pool for melanogenesis.
Clinical and Experimental Dermatology, Department of Biomedical Sciences, University of Bradford, West Yorkshire BD7 1DP, UK. k.schallreuter@bradford.ac.uk
Karin U
KU
Schallreuter
Umar
U
Wazir
Sonal
S
Kothari
Nicholas C J
NC
Gibbons
Jeremy
J
Moore
John M
JM
Wood
D002455:Cell Division; D004789:Enzyme Activation; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry* / Q000378:metabolism*; D008543:Melanins / Q000096:biosynthesis; D008544:Melanocytes / Q000378:metabolism* / Q000528:radiation effects; D008958:Models, Molecular; D010084:Oxidation-Reduction; D010649:Phenylalanine / Q000737:chemistry / Q000378:metabolism; D010651:Phenylalanine Hydroxylase / Q000737:chemistry* / Q000378:metabolism* / Q000528:radiation effects; D011487:Protein Conformation; D014443:Tyrosine / Q000737:chemistry* / Q000378:metabolism*; D014466:Ultraviolet Rays
D008543:Melanins; D014443:Tyrosine; D010649:Phenylalanine; D006861:Hydrogen Peroxide; D010651:Phenylalanine Hydroxylase