Bombesin

Department of Molecular and Integrative Physiology University of Michigan, Ann Arbor MI 48109
jawillms@med.umich.edu

Entry Version: 

Version 1.0, March 27, 2015

Citation: 

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Gene symbol:  GRP

1. General Information

Peptides of the bombesin family were first isolated from frog skin in the early 1970s.  Bombesin,  a 14 amino acid peptide was isolated from the skin of the amphibian Bombina bombina (Figure 1).  


Figure 1. Bombina bombina also known as the European fire-bellied toad. Image from en.wikipedia.org.

Its amino acid sequence was determined and it was shown to contain blocked amino and carboxyl terminals (1). Related peptides termed alytensin, ranatensin and litorin were isolated from other frogs (10).  Extracts from this and related frogs (Bombina variegate) had shown a number of actions on vascular smooth muscle, gall bladder contraction, gastric and pancreatic secretion, uterine smooth muscle and renal function.  Bombesin is the most potent known stimulator of gastrin release in dogs and humans and thereby stimulates gastric acid secretion (4).  It also stimulates CCK release (11).   

Using cDNA cloning the bombesin precursor was shown to contain 107 amino acids (44).  The precursor contained a signal peptide sequence and one copy of bombesin followed by a typical processing site. Antibodies against bombesin stained cells in the mammalian GI tract suggesting a counterpart (41). The first mammalian bombesin like peptide was isolated from pig gastric tissue and named gastrin-releasing peptide (GRP) because it had a potent action to release gastrin (34).  GRP is a 27 amino acid peptide and results from a 148 amino acid precursor (46).  Other subsequently identified related mammalian molecules include Neuromedin B  isolated from porcine spinal cord (35).  Neuromedin C was also isolated from pig spinal cord (36) but is now known to be GRP 18-27 (22).   All share a common carboxyl terminal sequence as shown in Table 1.

For a review of early work purifying mammalian bombesin-like peptides see (51).

The bombesin/GRP/NMC receptor was first characterized by the high affinity binding of radioiodinated bombesin.  The receptor was cloned from murine Swiss 3T3 cells (3, 45) and a distinct NMB receptor from rat esophagus (50).  Analysis of the predicted amino acid sequence revealed 7 putative transmembrane domains and the receptors were latter shown to be G-protein coupled.  Subsequently two other receptors, BRS-3 and BBE were identified (22, 39).  These four receptors are broadly distributed especially in the GI tract and brain.  The major form in the pancreas is the GRP-R also known as the BB2 receptor (39).  A number of bombesin receptor antagonists have also been developed most of which are modified bombesin peptides some having a reduced peptide bond (8, 16, 49).  One of the most potent is [D-Phe6]BN-(6-13) ethyl ester with a Ki of 5 nM to inhibit pancreatic amylase secretion stimulated by bombesin.  Bombesin peptide agonists and antagonists have also been proposed as imaging tools or to deliver chemotherapeutic agents to tumors with bombesin receptors.

2.  Bombesin and the Pancreas

Early studies indicated that bombesin administration that stimulated pancreatic secretion, also stimulated gastrin, CCK, and acetylcholine  release (2, 11, 33).  The secretion induced was enzyme rich and poor in bicarbonate.  Thus, an initial question was whether bombesin stimulated the pancreas directly or through a neural or hormonal intermediate. However, studies using antrectomized dogs to remove gastrin, CCK antagonists and atropine have shown in multiple species including humans that the effect of bombesin was primarily direct (17, 18, 29).  This was confirmed by the finding of specific bombesin receptors on isolated pancreatic acini and pancreatic AR42J cells (23, 30).  In vivo studies also reported a trophic effect on the exocrine pancreas although not as big as the response to caerulein (48).

Even after the discovery of mammalian GRP, bombesin continued in common use for the study of cellular mechanisms using isolated acini.  Bombesin and related peptides showed a slight biphasic dose response with reduced ability to stimulate in vitro amylase release from isolated guinea pig acinar cells at supramaximal concentrations; maximal secretion was observed at 100 nM (32, 47).   Similar results but with maximal secretion at 100 pM were seen in mouse acini (21).  By contrast in rat acini, bombesin induced a monophasic dose response with a maximal response at 1 nM (31, 38).   Bombesin activated IP3 and diacylglycerol production and intracellular calcium release (9, 31, 43).  Thus bombesin activates signaling pathways through heterotrimeric Gq/11 protein similar to CCK although by different receptors.  Other actions of bombesin shared with CCK have included activation of protein tyrosine kinases, PKC, phospholipase A2, p125FAK, ERK, JNK, p70 S6K and downregulation of c-Met (5, 7, 19, 24, 38).  Other differences between bombesin and CCK on acini are that bombesin induces less damage and ER stress (27).  In some studies bombesin is used as an alternative agonist to show that an event is not initiated by a single receptor (6).  Other studies have looked at the properties of bombesin receptors to internalize bombesin (54), their ability to induce residual stimulation (21), to desensitize (31), and to be regulated by CCK (53).

Bombesin by itself does not induce experimental pancreatitis in vivo or acinar cell damage in vitro similar to what is produced by caerulein (42,52).  Bombesin increases the activation of intracellular trypsin and the processing of procarboxypeptidase A1 in isolated acini (14).  However, the activated enzyme was secreted from the cell following bombesin stimulation thus plausibly explaining the lack of cell damage (14).  By contrast, when bombesin stimulation was combined with pancreatic duct obstruction, retention of active enzymes and pancreatitis resulted (40).  Another difference from caerulein is that bombesin failed to activate NF-κB (15).

Endogenenous GRP, the mamammalian equivalent of bombesin in the pancreas is primarily located in neurons.  Immunoflourescence has localized it to both pancreatic ganglia and in beaded neurons running between acini (13, 25, 37). The pig however, was the only species to have a high concentration of immunoreative GRP in the pancreas (37). Using rat pancreatic lobules, GRP was shown to stimulate acetylcholine release and about half of the effect on amylase release was blocked by neuronal or ganglionic blockers (12).  This suggests GRP may act both on neurons and acinar cells.  Studies in the pig have shown that electrical stimulation of the vagus releases GRP and its active fragment, GRP (18-27) both in vivo and in the perfused pancreas (25, 26).  Furthermore bombesin receptor antagonists inhibited secretion induced by vagal stimulation by 33% (20).  Thus in the pig, GRP plays a role in endogenous pancreatic secretion although the role in other species may be less.

3.  Tools to study bombesin

a. Synthetic Peptide

Bombesin can be obtained from multiple sources including Sogma-Aldrich, Research Plus, Anaspec and Abbiotec.

b. Antibodies

Antibodies against bombesin are available from Genway and Phoenix Pharmaceuticals. 

c. ELISA and RIA

An RIA kit for GRP is available from Phoenix Pharmaceuticals.  An ELISA kit against Bombesin is available from MyBioSource and against mouse GRP from Antibodies-online.

d. Antagonists

Peptide antagonists against the Bombesin receptor are available from Sigma-Aldrich and Tocris

4. References

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  2. Basso N, Giri S, Improta G, et al. External pancreatic secretion after bombesin infusion in man. Gut 16(12):994-998, 1975. PMID: 1218823.
  3. Battey JF, Way JM, Corjay MH, et al. Molecular cloning of the bombesin/gastrin-releasing peptide receptor from Swiss 3T3 cells. Proc Natl Acad Sci USA 88(2):395-399, 1991. PMID: 1671171.
  4. Bertaccini G, Erspamer V, Melchiorri P, Sopranzi N. Gastrin release by bombesin in the dog. Br J Pharmacol 52(2):219-225, 1974. PMID: 4451816.
  5. Bragado MJ, Groblewski GE, Williams JA. p70s6k is activated by CCK in rat pancreatic acini. Am J Physiol 273:C101-C109, 1997. PMID: 9252447.
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  9. Deschodt-lanckman M, Robberecht P, De neef P, Lammens M, Christophe J. In vitro action of bombesin and bombesin-like peptides on amylase secretion, calcium efflux, and adenylate cyclase activity in the rat pancreas: a comparison with other secretagogues. J Clin Invest 58(4):891-898, 1976. PMID: 184111.
  10. Erspamer, V. Discovery, Isolation, and Characterization of Bombesin-like Peptides. Ann N Y Acad Sci 547: 3–9, 1988. PMID: 3071223.
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  17. Herzig KH, Louie DS, Owyang C. In vivo action of bombesin on exocrine pancreatic secretion in the rat: independent of cholecystokinin and cholinergic mediation. Pancreas 3(3):292-296, 1988. PMID: 3387423.
  18. Hildebrand P, Drewe J, Luo H, Ketterer S, Gyr K, Beglinger C. Role of cholecystokinin in mediating GRP-stimulated gastric, biliary and pancreatic functions in man. Regul Pept 41(2):119-129, 1992. PMID: 1438984.
  19. Hoffmann KM, Tapia JA, Berna MJ, et al. Gastrointestinal hormones cause rapid c-Met receptor down-regulation by a novel mechanism involving clathrin-mediated endocytosis and a lysosome-dependent mechanism. J Biol Chem 281(49):37705-37719, 2006. PMID: 17035232.
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  24. Kiehne K, Herzig KH, Fölsch UR. Differential activation of p42ERK2 and p125FAK by cholecystokinin and bombesin in the secretion and proliferation of the pancreatic amphicrine cell line AR42J. Pancreatology 2(1):46-53, 2002. PMID: 12120007.
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  27. Kubisch CH, Logsdon CD. Secretagogues differentially activate endoplasmic reticulum stress responses in pancreatic acinar cells. Am J Physiol Gastrointest Liver Physiol 292(6):G1804-G1812, 2007. PMID: 17431218.
  28. Lee PC, Jensen RT, Gardner JD. Bombesin-induced desensitization of enzyme secretion in dispersed acini from guinea pig pancreas. Am J Physiol 238(3):G213-G218, 1980. PMID: 6154421.
  29. Liehr RM, Reidelberger RD, Varga G, Solomon TE. Mechanism of bombesin-induced pancreatic secretion in unanesthetized rats. Peptides. 14(4): 717-723, 1993. PMID: 8234015.
  30. Logsdon CD, Zhang JC, Guthrie J, Vigna S, Williams JA. Bombesin binding and biological effects on pancreatic acinar AR42J cells. Biochem Biophys Res Commun 144(1):463-468, 1987. PMID: 2437913.
  31. Matozaki T, Zhu WY, Tsunoda Y, Göke B, Williams JA. Intracellular mediators of bombesin action on rat pancreatic acinar cells. Am J Physiol 260: G858-G864, 1991. PMID: 1711779.
  32. May RJ, Conlon TP, Erspamer V, Gardner JD. Actions of peptides isolated from amphibian skin on pancreatic acinar cells. Am J Physiol 235: E112-E118, 1978. PMID: 210673.
  33. Mcdonald TJ, Ghatei MA, Bloom SR, et al. Dose-response comparisons of canine plasma gastroenteropancreatic hormone responses to bombesin and the porcine gastrin-releasing peptide (GRP). Regul Pept 5(2):125-137, 1983. PMID: 6338565.
  34. Mcdonald TJ, Jörnvall H, Nilsson G, et al. Characterization of a gastrin releasing peptide from porcine non-antral gastric tissue. Biochem Biophys Res Commun 90(1):227-233, 1979. PMID: 496973.
  35. Minamino N, Kangawa K, Matsuo H. Neuromedin B: a novel bombesin-like peptide identified in porcine spinal cord. Biochem Biophys Res Commun 114(2):541-548, 1983. PMID: 6882442.
  36. Minamino N, Kangawa K, Matsuo H. Neuromedin C: a bombesin-like peptide identified in porcine spinal cord. Biochem Biophys Res Commun 119(1):14-20, 1984. PMID: 6546686.
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  38. Nishino H, Tsunoda Y, Owyang C. Mammalian bombesin receptors are coupled to multiple signal transduction pathways in pancreatic acini. Am J Physiol 274: G525-G534, 1998. PMID: 9530154.
  39. Ohki-Hamazaki H, Iwabuchi M, Maekawa F. Development and function of bombesin-like peptides and their receptors. Int J Dev Biol 49(2-3):293-300, 2005. PMID: 15906244.
  40. Otani T, Matsukura A, Takamoto T, et al. Effects of pancreatic duct ligation on pancreatic response to bombesin. Am J Physiol Gastrointest Liver Physiol 290(4):G633-G639, 2006.
  41. Polak JM, Bloom SR, Hobbs S, Solcia E, Pearse AG. Distribution of a bombesin-like peptide in human gastrointestinal tract. Lancet 1(7969):1109-1110, 1976. PMID: 57512.
  42. Powers RE, Grady T, Orchard JL, Gilrane TB. Different effects of hyperstimulation by similar classes of secretagogues on the exocrine pancreas. Pancreas 8(1):58-63, 1993.
  43. Pralong WF, Wollheim CB, Bruzzone R. Measurement of cytosolic free Ca2+ in individual pancreatic acini. FEBS Lett 242(1):79-84, 1988. PMID: 2462514.
  44. Richter K, Egger R, Kreil G. Molecular cloning of a cDNA encoding the bombesin precursor in skin of Bombina variegata. FEBS Lett 262(2):353-355, 1990. PMID: 2335218.
  45. Spindel ER, Giladi E, Brehm P, Goodman RH, Segerson TP. Cloning and functional characterization of a complementary DNA encoding the murine fibroblast bombesin/gastrin-releasing peptide receptor. Mol Endocrinol 4(12):1956-1963, 1990. PMID: 1707129.
  46. Spindel ER, Chin WW, Price J, Rees LH, Besser GM, Habener JF. Cloning and characterization of cDNAs encoding human gastrin-releasing peptide. Proc Natl Acad Sci USA 81(18):5699-5703, 1984. PMID: 6207529.
  47. Uhlemann ER, Rottman AJ, Gardner JD. Actions of peptides isolated from amphibian skin on amylase release from dispersed pancreatic acini. Am J Physiol 236(5):E571-E576, 1979. PMID: 443378.
  48. Varga G, Papp M, Dobronyi I, Scarpignato C. Effect of bombesin and its mammalian counterpart, GRP, on exocrine pancreas in the rat. Digestion 41(4):229-236, 1988. PMID: 2468544.
  49. Von Schrenck T, Wang LH, Coy DH, Villanueva ML, Mantey S, Jensen RT. Potent bombesin receptor antagonists distinguish receptor subtypes. Am J Physiol 259: G468-G473, 1990. PMID: 2169207.
  50. Wada E, Way J, Shapira H, et al. cDNA cloning, characterization, and brain region-specific expression of a neuromedin-B-preferring bombesin receptor. Neuron 6(3):421-430. 1991. PMID: 1848080
  51. Walsh JH, Wong HC, Dockray GJ. Bombesin-like peptides in mammals. Fed Proc 38(9): 2315-2319, 1979. PMID: 456618.
  52. Wisner JR, Ozawa S, Renner IG. Pancreatic exocrine function in unconscious rats treated with submaximal, maximal, and supramaximal doses of bombesin tetradecapeptide. Pancreas 4(1):83-9, 1989. PMID: 2717605.
  53. Younes M, Wank SA, Vinayek R, Jensen RT, Gardner JD. Regulation of bombesin receptors on pancreatic acini by cholecystokinin. Am J Physiol 256: G291-G298, 1989. PMID: 2465695.
  54. Zhu WY, Göke B, Williams JA. Binding, internalization, and processing of bombesin by rat pancreatic acini. Am J Physiol 261: G57-G64, 1991. PMID: 1650142.