geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Direct identification of a second distinct site of contact between cholecystokinin and its receptor


, : Direct identification of a second distinct site of contact between cholecystokinin and its receptor. Journal of Biological Chemistry 273(21): 12988-12993

We have developed a biologically active analogue of cholecystokinin (CCK) that incorporates a photolabile benzoylphenylalanine (Bpa) moiety in the middle of its pharmacophoric domain, which efficiently establishes a covalent bond with an interacting domain of the CCK receptor. This probe incorporated L-Bpa in the position of Gly29 of the well characterized, radioiodinatable CCK analogue, D-Tyr-Gly-[(Nle28,31)CCK-26-33]. It was a potent pancreatic secretagogue (EC50 = 28 +/- 6 nM) that was equally efficacious with natural CCK, and bound to the CCK receptor with moderate affinity (IC50 = 450 +/- 126 nM). This was adequate to allow specific covalent labeling of the receptor. The labeled domain was within the cyanogen bromide fragment of the receptor including the top of TM6 (the sixth transmembrane domain), the third extracellular loop, and TM7 (the seventh transmembrane domain), as proven by direct Edman degradation sequencing. When this fragment was modified by the replacement of Val342 with Met to generate an additional site of cyanogen bromide cleavage, the labeled fragment was reduced in apparent size consistent with its representing the carboxyl-terminal portion of this fragment. Radiochemical sequencing of that fragment demonstrated covalent attachment of the probe to His347 and Leu348 in this domain. This represents the second experimentally demonstrated contact between a CCK analogue and this receptor, complementing the labeling of the domain just above TM1 (the first transmembrane domain) by a photolabile residue at the carboxyl terminus of CCK (Ji, Z. S., Hadac, E. M., Henne, R. M., Patel, S. A., Lybrand, T. P., and Miller, L. J. (1997) J. Biol. Chem. 272, 24393-24401). Both contacts are consistent with the conformational model of CCK binding proposed on the basis of the initial contact.

US$19.90

PMID: 9582333

DOI: 10.1074/jbc.273.21.12988


Other references

Abe S.; Kuramitsu I.; Yonahara Y., 1967: Studies on the inst radio isotope renogram in juvenile hypertension abstract human kidney disease. Kaku Igaku 4(3/4):

Theodorides, J., 1980: Hotes nouveaux de Gregarina joliveti (Theod. 1958) au Maroc. Annales de Parasitologie Humaine et Comparee 55(2): 247-248

Thomas, T.T., 1911: VII. Laceration of the Axillary Portion of the Capsule of the Shoulder-Joint as a Factor in the Etiology of Traumatic Combined Paralysis of the Upper Extremity. Annals of Surgery 53(1): 77-105

Konteatis, Z.; Siciliano, S.J.; Springer, M.S., 1997: Assay to identify human C5A antagonists and agonists. Official Gazette of the United States Patent & Trademark Office Patents 1196(4): 2640, March 25

Fennewald, S.M.; Rando, R.F., 1995: Inhibition of high affinity basic fibroblast growth factor binding by oligonucleotides. Oligonucleotides can be used to inhibit the binding of basic fibroblast growth factor to cells. Though standard phosphodiester oligonucleotides show a slight inhibition of binding, the oligonucleotides with phosphorothioate internucleoside linkage...

Smilie, Jl, 1976: An adventure in energy! Use the sun to help heat your house--block the sun rays for cooling. Coop ext pub: 976, 1875

Chichester, F.; Smith, S., 1979: Glasshouse evaluation of forage grass nutrient responses on lignite mine site overburden material from east central Texas. Agronomy abstracts: 69

Zhang, H.; Cao, M.; Wu, W.; Xu, H.; Cheng, S.; Fan, L-Juan., 2014: Polyacrylonitrile/noble metal/SiO₂ nanofibers as substrates for the amplified detection of picomolar amounts of metal ions through plasmon-enhanced fluorescence. Electrospun polymer/noble metal hybrid nanofibers have developed rapidly as surface-enhanced Raman scattering (SERS)-active substrates over the last few years. However, polymer/noble metal nanofibers with plasmon-enhanced fluorescence (PEF) activi...

Saketkoo, L.Ann.; Escorpizo, R.; Keen, K.J.; Fligelstone, K.; Distler, O., 2013: International Classification of Functioning, Disability and Health Core Set construction in systemic sclerosis and other rheumatic diseases: a EUSTAR initiative. To outline rationale and potential strategies for rheumatology experts to be able to develop disease-specific Core Sets under the framework of the International Classification of Functioning, Disability and Health (ICF). ICF is a universal framewo...

Nykänen, M.; Vehviläinen-Julkunen, K.; Klemetti, R., 2017: The expectations of antenatal screening and experiences of the first-trimester screening scan. to describe the expectations to and knowledge of antenatal screening of expecting parents, and their experiences concerning the first-trimester screening scan. survey study with a consecutive sampling method. two hospital districts in Finland, nam...