Open AccessA human melanoma‐derived cell line (IGR39) with a very high number of vasoactive‐intestinal‐peptide (VIP) receptors
Author(s) -
LUIS José,
MARTIN JeanMichel,
BATTARI Assou,
REYNIER Max,
MARVALDI Jacques,
PICHON Jacques
Publication year - 1989
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1989.tb14664.x
Subject(s) - vasoactive intestinal peptide , secretin , secretin family , peptide , receptor , chemistry , dissociation constant , binding site , biochemistry , peptide hormone , affinity chromatography , biology , neuropeptide , enzyme , secretion
Using mono[ 125 I]iodinated vasoactive intestinal peptide ( 125 I‐VIP), a very high number of specific binding sites for VIP were identified at the surface of the human melanoma cell line IGR39. The Scatchard analysis of competitive displacement experiments between native VIP and 125 I‐VIP was consistent with the existence of two classes of VIP‐binding sites. IGR39 cells possess 0.54 × 10 6 high‐affinity sites with a dissociation constant ( K d ) of 0.66 nM and 1.3 × 10 6 sites of moderate affinity with a K d of 4.7 nM. Pharmacological studies indicated that the order of potency in inhibiting 125 I‐VIP binding of the VIP secretin family peptides was VIP Gt; peptide histidine methioninamide > human growth‐hormone‐releasing factor(1–44) > secretin. Glucagon has no effect on the binding of the labelled peptide. By means of photoaffinity labelling a polypeptide of M r 63000 was characterized. The labelling of this species was completely abolished by native VIP. The order of potency of VIP‐related peptides in inhibiting 125 I‐VIP cross‐linking to its receptor was the same as in the competition experiments. The glycoprotein nature of the VIP‐binding sites of IGR39 cells has been investigated by affinity chromatography on wheat‐germ‐agglutinin–Sepharose.