Identification of the F1F0 mitochondrial ATPase as a target for modulating skin pigmentation by screening a tagged triazine library in zebrafish.
A triazine-based combinatorial library of small molecules was screened in zebrafish to identify compounds that produced interesting phenotypes. One compound (of 1536 screened) induced a dramatic increase in the pigmentation of early stage zebrafish embryos. This compound, PPA, was also found to increase pigmentation in cultured mammalian melanocytes. The cellular target was identified as the mitochondrial F1F0-ATP synthase (ATPase) by affinity chromatography. Oligomycin, a small molecule known to inhibit the mitochondrial ATPase, competed with PPA for its cellular target in melanocytes. In addition, PPA was shown to alter the membrane potential of mitochondria, consistent with inhibition of the mitochondrial ATPase. Thus, PPA has been successfully used as a chemical probe in a forward chemical genetic approach to establish a link between the phenotype and the protein. The results attest to the power of screening small molecule libraries in zebrafish as a means of identifying mammalian targets and suggest the mitochondrial ATPase as a target for modulating pigmentation in both melanocytes and melanoma cells.
Jung
Da-Woon
DW
Department of Chemistry, New York University, New York, NY 10003, USA.
Williams
Darren
D
Khersonsky
Sonya M
SM
Kang
Tae-Wook
TW
Heidary
Noushin
N
Chang
Young-Tae
YT
Orlow
Seth J
SJ
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D004625:Embryo, Nonmammalian; D008543:Melanins / Q000378:metabolism; D008544:Melanocytes / Q000502:physiology; D008564:Membrane Potentials; D051379:Mice; D008810:Mice, Inbred C57BL; D051336:Mitochondrial Membranes / Q000502:physiology; D024101:Mitochondrial Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*; D025261:Mitochondrial Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000502:physiology*; D008969:Molecular Sequence Data; D009840:Oligomycins / Q000494:pharmacology; D012880:Skin Pigmentation / Q000502:physiology*; D014227:Triazines / Q000494:pharmacology; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000037:antagonists & inhibitors / Q000502:physiology*
D008543:Melanins; D024101:Mitochondrial Proteins; D009840:Oligomycins; D014227:Triazines; D029961:Zebrafish Proteins; C474451:F1F0-ATP synthase; D025261:Mitochondrial Proton-Translocating ATPases