Structurally distinctive vasoactive intestinal peptides from rat basophilic leukemia cells.
Peptides recognized by rabbit antibodies to vasoactive intestinal peptide (VIP) were extracted from diisopropyl fluorophosphate-treated rat basophilic leukemia (RBL) cells and resolved by filtration on Sephadex G-25 in 50 mM acetic acid. The immunoreactive VIPs of RBL cells eluted from Sephadex G-25 at 35-41%, 53-60%, and 69-73% bed volume, but not at 63-68% as for the neuropeptide VIP1-28. The two forms of immunoreactive VIP larger than VIP1-28 reacted with antibodies to both VIP1-9 and VIP10-28, but the smallest was bound only by antibodies to VIP10-28. The smallest immunoreactive VIP was purified by ion-exchange and reverse-phase high-performance liquid chromatography, and the amino acid sequence was determined to be that of VIP10-28 with asparagine-free acid at the carboxyl terminus rather than the amide of VIP neuropeptide. Challenge of RBL cells with 1 microM ionophore A23187 at 37 degrees C released VIP10-28 rapidly to a mean of 75% at 5 min and 77% at 30 min. The VIP generated and released by mast cells thus consists of a mixture of peptides that all differ structurally from the neuropeptide VIP.
Goetzl
E J
EJ
Howard Hughes Medical Institute, University of California Medical Center, San Francisco 94143-0724.
Sreedharan
S P
SP
Turck
C W
CW
D000595:Amino Acid Sequence; D000818:Animals; D001491:Basophils / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D007420:Intestine, Large / Q000032:analysis; D007421:Intestine, Small / Q000032:analysis; D007942:Leukemia, Experimental / Q000378:metabolism*; D008407:Mast Cells / Q000032:analysis; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011863:Radioimmunoassay; D051381:Rats; D011919:Rats, Inbred Strains; D014660:Vasoactive Intestinal Peptide / Q000276:immunology / Q000302:isolation & purification*
D014660:Vasoactive Intestinal Peptide