Functional characterization of L-tryptophan transport across mammalian cornea
Keywords:
Cornea, Amino acid, Tryptophan, TransportAbstract
In last few years transporter targeted drug delivery has drawn attention of research to identify and explore various nutrient transport systems including amino acid transporters for better drug delivery. The aim of present research work is to investigate the transport characteristics of L-tryptophan (L-try) across goat cornea. Transport of L-try was investigated using a glass diffusion cell for effect of concentration, pH, presence of other amino acids or metabolic inhibitor or dipeptide and tripeptide. The amount of L-try transported increased as the pH of L-try aqueous solution increased from 5 to 9. Inhibition was observed in L-try transport in absence of sodium ions where L-try solution was made isotonic with dextrose. Amino acids like L-histidine, L-arginine, L-lysine (cationic), L-glutamic acid, L-aspartic acid (anionic), glycine and L-proline (neutral) inhibited the L-try transport as compared to control (L-try alone). In presence of sodium azide and Ouabain the inhibition in L-try transport across goat cornea was observed while no marked inhibition was observed on L-try transport across goat cornea in presence of aspartame and glutathione. The L-try transport was favored up to concentration 1% w/v and at higher pH in presence of sodium ions through excised goat cornea. Functional presence of a sodium dependent L-try transport system as inhibited by ouabain having affinity to cationic and neutral amino acid is evident on goat cornea.
References
Christensen H.N. Role of amino acid transport
and counter transport in nutrition and metabolism.
Physiol Rev. 1990;70:43-77.
Christensen H.N., Albritton L.M., Kakuda D.K.
Gene-product designations for amino acid
transporters. J Exp Biol. 1994;196:51-57.
Broer S., Wagner C.A., Lang F. Function and
structure of heterodimeric amino acid
transporters. Am J Physiol. 2001;281:C1077-
C109.
Palacin M., Estevez R., Bertran J. Molecular
biology of mammalian plasma membrane amino
acid transporters. Physiol Rev. 1998;78:969-1054.
Deves R., Boyd C.A. Transporters for cationic
amino acids in animal cells: discovery, structure,
and function. Physiol Rev. 1998;78:487-545.
Kekuda R., Torres Z.V., Fei Y.J. Molecular and
functional characterization of intestinal Na+-
dependent neutral amino acid transporter B0. Am
J Physiol.1997;272:G1463-G1472.
Torrents D., Estevez R., Pineda M. Identification
and characterization of a membrane protein (y+L
amino acid transporter-1) that associates with
F2hc to encode the amino acid transport activity
y
+L: a candidate gene for lysinuric protein
intolerance. J Biol Chem. 1998;273:32437-32445.
Pfeiffer R., Rossier G., Spindler B. Amino acid
transport of y+L-type by heterodimers of
F2hc/CD98 and members of the glycoproteinassociated amino acid transporter family. EMBO
J. 1999;18:49-57.
Anand B.S., Mitra A.K. Mechanism of corneal
permeation of L-valyl ester of acyclovir: targeting
the oligopeptide transporter on the rabbit cornea.
Pharm Res. 2002;19:1194-1202.
Jain V.B., Pal D.,Gunda S.,Nashed Y.,Ganapathy
V., Mitra A.K. identification of a Na+-dependent
cationic and neutral amino acid transporter,B 0,+ in
human and rabbit cornea. Mol Pharm.
;1:338-346.
Anand B.S., Katragadda S., Nashed Y.E., Mitra
A.K. Amino acid prodrugs of acyclovir as
possible antiviral agents against ocular HSV-1
infections: interactions with the neutral and
cationic amino acid transporter on the corneal
epithelium. Curr Eye Res. 2004;29:153-66.;
Majumdar S., Nashed Y.E, Patel K., Jain R.,
Itahashi M., Neumann D.M., Hill J.M., Mitra
A.K. Dipeptide monoester ganciclovir prodrugs
for treating HSV-1-induced corneal epithelial and
stromal keratitis: in vitro and in vivo evaluations.
J Ocul Pharmacol Ther. 2005;21:463-74.
Nielsen C.U., Vabeno J., Andersen R., Brodin B.,
Steffansen B. Recent advances in therapeutic
applications of human peptide transporters. Expert
Opin Ther Patents. 2005;15:153–166.
Niederkorn J.Y., Meyer D.R., Ubelaker J.E.,
Martin J.H. Ultrastructural and
immunohistological characterization of the SIRC
corneal cell line. In Vitro Cell Dev Biol. 1990;
:923–930.
Carl G.F., Hoffman W.H., Blankenship P.R.,
Litaker M.S., Hoffman M.G., Mabe P.A. Diabetic
Ketoacidosis depletes plasma tryptophan. Endocr
Res. 2002;28:91-102.
Vrensen G.F., Van M.J., Jonges R., Voorhout W.,
Breipohl W., Wegener A.R. Tryptophan
deficiency arrests chromatin breakdown in
secondary lens fibers of rats. Exp Eye Res.
;78:661-672.
Rathore M.S., Majumdar D.K. Effect of
Formulation Factors on In Vitro Transcorneal
Permeation of Gatifloxacin From Aqueous Drops.
AAPS PharmSciTech. 2006; 7: E1-E6.