Specific recognition of the human neuroendocrine receptor for vasoactive intestinal peptide by anti-peptide antibodies

Molecular and Cellular Neurosciences
E J GoetzlS P Sreedharan

Abstract

Rabbit polyclonal IgG antibodies were generated to three distinct synthetic peptide substituents of the human neuroendocrine-type 7 transmembrane-domain receptor for vasoactive intestinal peptide (VIP), including a portion of the amino-terminus, first extracellular loop, and carboxyl-terminus. Immunofluorescent staining of both human K293 cell transfectants, expressing recombinant VIP receptors, and HT-29 human intestinal epithelial cells, bearing native VIP receptors, was observed with each of the antibodies and was eliminated specifically after absorption of antibodies with the respective peptide immunogen. Each of the antibodies recognized the same approximately 70-kDa membrane proteins, extracted from both K293 cell transfectants and HT-29 cells, in sodium dodecyl sulfate-polyacrylamide gel electrophoresis blots. Neither IgG nor Fab preparations of the antibodies inhibited VIP binding to cellular receptors at a concentration of 1 microgram/ml, that yielded optimal immunofluorescence, or at 5-300 micrograms/ml. In contrast, 5-200 micrograms/ml of anti-peptide antibodies as IgG, but not Fab, significantly inhibited the increase in concentration of cyclic AMP in HT-29 cells elicited by 1 nM VIP, without affecting the greater i...Continue Reading

Citations

Nov 8, 2001·Proceedings of the National Academy of Sciences of the United States of America·E J GoetzlA J Harmar
Nov 2, 2001·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·J K VoiceE J Goetzl
Jan 25, 2003·Journal of Gastroenterology and Hepatology·Akihiko ItohTadao Bamba
May 20, 2006·The Journal of Immunology : Official Journal of the American Association of Immunologists·Mei-Chuan HuangEdward J Goetzl
Feb 24, 2001·Annals of the New York Academy of Sciences·R BustoJ C Prieto
Jul 17, 1999·The American Journal of Physiology·R BustoJ C Prieto

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