Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus.

Authors

William Grosvenor, Monell Chemical Senses Center, Philadelphia, PA 19104-3308, USAFollow
Yuri Kaulin, Monell Chemical Senses Center, Philadelphia, PA 19104-3308, USA, Institute of Cytology, Russian Academy of Sciences, St. Petersburg 194064, Russia Monell Chemical Senses Center, Philadelphia, PA 19104-3308, USA, Institute of Cytology, Russian Academy of Sciences, St. Petersburg 194064, Russia, Department of Pathology, Anatomy & Cell Biology; Thomas Jefferson University; Philadelphia, PA 19107-6799Follow
Andrew I Spielman, New York University College of Dentistry, New York, NY 10010, USAFollow
Douglas L Bayley, Monell Chemical Senses Center, Philadelphia, PA 19104-3308, USAFollow
D Lynn Kalinoski, Monell Chemical Senses Center, Philadelphia, PA 19104-3308, USA, UCSD Thornton Hospital, San Diego, CA 92037Follow
John H Teeter, Monell Chemical Senses Center, Philadelphia, PA 19104-3308, USA, Institute of Neurological Sciences, University of Pennsylvania, Philadelphia, PA 19104, USAFollow
Joseph G Brand, Monell Chemical Senses Center, Philadelphia, PA 19104-3308, USA, Veterans Affairs Medical Center, Philadelphia, PA 19104, USA, Department of Biochemistry, School of Dental Medicine, University of Pennsylvania, Philadelphia, PA 19104, USAFollow

Document Type

Article

Publication Date

7-28-2004

Comments

This article has been peer reviewed and is published in BMC Neuroscience Volume 5, 28 July 2004, Article Number 25. The published version is available at DOI: 10.1186/1471-2202-5-25. Copyright © BioMed Central Ltd.

Abstract

BACKGROUND: The channel catfish, Ictalurus punctatus, is invested with a high density of cutaneous taste receptors, particularly on the barbel appendages. Many of these receptors are sensitive to selected amino acids, one of these being a receptor for L-arginine (L-Arg). Previous neurophysiological and biophysical studies suggested that this taste receptor is coupled directly to a cation channel and behaves as a ligand-gated ion channel receptor (LGICR). Earlier studies demonstrated that two lectins, Ricinus communis agglutinin I (RCA-I) and Phaseolus vulgaris Erythroagglutinin (PHA-E), inhibited the binding of L-Arg to its presumed receptor sites, and that PHA-E inhibited the L-Arg-stimulated ion conductance of barbel membranes reconstituted into lipid bilayers. RESULTS: Both PHA-E and RCA-I almost exclusively labeled an 82-84 kDa protein band of an SDS-PAGE of solubilized barbel taste epithelial membranes. Further, both rhodamine-conjugated RCA-I and polyclonal antibodies raised to the 82-84 kDa electroeluted peptides labeled the apical region of catfish taste buds. Because of the specificity shown by RCA-I, lectin affinity was chosen as the first of a three-step procedure designed to enrich the presumed LGICR for L-Arg. Purified and CHAPS-solubilized taste epithelial membrane proteins were subjected successively to (1), lectin (RCA-I) affinity; (2), gel filtration (Sephacryl S-300HR); and (3), ion exchange chromatography. All fractions from each chromatography step were evaluated for L-Arg-induced ion channel activity by reconstituting each fraction into a lipid bilayer. Active fractions demonstrated L-Arg-induced channel activity that was inhibited by D-arginine (D-Arg) with kinetics nearly identical to those reported earlier for L-Arg-stimulated ion channels of native barbel membranes reconstituted into lipid bilayers. After the final enrichment step, SDS-PAGE of the active ion channel protein fraction revealed a single band at 82-84 kDa which may be interpreted as a component of a multimeric receptor/channel complex. CONCLUSIONS: The data are consistent with the supposition that the L-Arg receptor is a LGICR. This taste receptor remains active during biochemical enrichment procedures. This is the first report of enrichment of an active LGICR from the taste system of vertebrata.

Recommended Citation

Grosvenor, William; Kaulin, Yuri; Spielman, Andrew I; Bayley, Douglas L; Kalinoski, D Lynn; Teeter, John H; and Brand, Joseph G, "Biochemical enrichment and biophysical characterization of a taste receptor for L-arginine from the catfish, Ictalurus puntatus." (2004). Department of Pathology, Anatomy, and Cell Biology Faculty Papers. Paper 40.
https://jdc.jefferson.edu/pacbfp/40

PubMed ID

15282034