Tyrosine transport in a human melanoma cell line as a basis for selective transport of cytotoxic analogues.
Tyrosine is an essential amino acid for the initial step of melanin synthesis, yet little is known concerning its transport in melanocytes. As an important first step in the development of new anti-melanoma agents based upon chemical and pharmacological modifications of melanin synthesis, the present study characterized the transport mechanism of tyrosine in vitro using the human melanoma cell line SK-MEL 23. Several tyrosine transport systems may be involved in melanocytes: systems L and T, which transport neutral amino acids with branched or aromatic side chains, and systems A and ASC, which transport neutral amino acids with smaller side chains. In order to determine which system or combination of systems is involved in tyrosine transport in melanoma cells, studies of kinetics, Na(+)-dependence and competitive inhibition were undertaken. The Km and Vmax. for the Na(+)-independent transport system were found to be 0.164 +/- 0.016 mM and 21.6 +/- 1.1 nmol/min per mg of protein respectively. This transport was preferentially inhibited by the system L specific analogue, 2-aminobicyclo[2.2.1]heptane-2-carboxylic acid, the system T substrate tryptophan, and the sulphur homologue of tyrosine, 4-S-cysteinylphenol. Sequential addition of these inhibitors at increasing concentrations indicated that they inhibit the same transporter. Our results suggest that tyrosine transport in SK-MEL 23 melanoma cells is similar to system L transport previously characterized in other cell types. This one transport system appears to supply all the tyrosine required for both cell growth and melanin synthesis. The transport system may be subject to manipulation by melanogenic stimulating factors, making the transport of cytotoxic tyrosine analogues an important area for further study.
Pankovich
J M
JM
Division of Dermatology and Cutaneous Sciences, University of Alberta, Edmonton, Canada.
Jimbow
K
K
D000596:Amino Acids / Q000378:metabolism; D000598:Amino Acids, Cyclic; D001693:Biological Transport, Active; D003545:Cysteine / Q000031:analogs & derivatives* / Q000378:metabolism; D016503:Drug Delivery Systems; D005069:Evaluation Studies as Topic; D006801:Humans; D007700:Kinetics; D008545:Melanoma / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000378:metabolism
D000596:Amino Acids; D000598:Amino Acids, Cyclic; C006504:2-aminobicyclo(2,2,1)heptane-2-carboxylic acid; D014443:Tyrosine; C043036:4-S-cysteinylphenol; D012964:Sodium; D003545:Cysteine