Abstract
Data concerning the pathophysiological role of extracellular S100A4, a member of the multigenic family of Ca2+-modulated S100 proteins, and its interaction with the receptor for advanced glycation endproducts (RAGE) or other putative receptors in tumorigenesis, metastasis, and inflammatory processes in vivo are scarce. One reason is the shortage of suitable radiotracer methods. We report a novel methodology using recombinant human S100A4 as potential probe for molecular imaging and functional characterization of this interaction. Therefore, human S100A4 was cloned as GST fusion protein in the bacterial expression vector pGEX-6P-1 and expressed in E. coli strain BL21. Purified recombinant human S100A4 was radiolabeled with the positron emitter fluorine-18 (18F) by conjugation with N-succinimidyl-4-[18F]fluorobenzoate ([18F]SFB). The radioligand [18F]fluorobenzoyl-S100A4 (18F-S100A4) was used in cell binding experiments in RAGE-bearing human melanoma cells and endothelial cells in vitro, and in both biodistribution experiments and small animal positron emission tomography (PET) studies in normal rats in vivo. The cellular association and tissue-specific distribution of 18F-S100A4 in vitro and in vivo correlated well with the protein expression and anatomical localization of RAGE, e.g., in the vascular system and in lung. Compared to other S100 RAGE radioligands, the overall findings of this study indicate that extracellular S100A4 in vivo shows only a moderate interaction with RAGE and, furthermore, exhibits a substantially faster metabolic degradation. On the other hand, the approach allows the use of quantitative small animal PET and provides a novel probe to both delineate functional expression and differentiate multiligand interaction of RAGE under normal and pathophysiological conditions in rodent models of disease.
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References
Abe R, Shimizu T, Sugawara H, Watanabe H, Nakamura H, Choei H, Sasaki N, Yamagishi S, Takeuchi M, Shimizu H (2004) Regulation of human melanoma growth and metastasis by AGE-AGE receptor interactions. J Invest Dermatol 122:461–467
Adachi H, Tsujimoto M (2006) Endothelial scavenger receptors. Prog Lipid Res 45:379–404
Arumugam T, Simeone DM, Schmidt AM, Logsdon CD (2004) S100P stimulates cell proliferation and survival via receptor for activated glycation end products (RAGE). J Biol Chem 279:5059–5065
Berge G, Maelandsmo GM (2010) Evaluation of potential interactions between the metastasis-associated protein S100A4 and the tumor suppressor protein p53. Amino Acids (in press)
Boyd JH, Kan B, Roberts H, Wang Y, Walley KR (2008) S100A8 and S100A9 mediate endotoxin-induced cardiomyocyte dysfunction via the receptor for advanced glycation end products. Circ Res 102:1239–1246
Boye K, Maelandsmo GM (2010) S100A4 and metastasis: a small actor playing many roles. Am J Pathol 176:528–535
Cunningham MF, Docherty NG, Burke JP, O’Connell PR (2010) S100A4 expression is increased in stricture fibroblasts from patients with fibrostenosing Crohn’s disease and promotes intestinal fibroblast migration. Am J Physiol Gastrointest Liver Physiol 299:G457–G466
Dattilo BM, Fritz G, Leclerc E, Kooi CW, Heizmann CW, Chazin WJ (2007) The extracellular region of the receptor for advanced glycation end products is composed of two independent structural units. Biochemistry 46:6957–6970
Donato R (2007) RAGE: a single receptor for several ligands and different cellular responses: the case of certain S100 proteins. Curr Mol Med 7:711–724
Fernandez-Fernandez MR, Veprintsev DB, Fersht AR (2005) Proteins of the S100 family regulate the oligomerization of p53 tumor suppressor. Proc Natl Acad Sci USA 102:4735–4740
Garrett SC, Varney KM, Weber DJ, Bresnick AR (2006) S100A4, a mediator of metastasis. J Biol Chem 281:677–680
Gebhardt C et al (2008) RAGE signaling sustains inflammation and promotes tumor development. J Exp Med 205:275–285
Ghavami S, Rashedi I, Dattilo BM, Eshraghi M, Chazin WJ, Hashemi M, Wesselborg S, Kerkhoff C, Los M (2008) S100A8/A9 at low concentration promotes tumor cell growth via RAGE ligation and MAP kinase-dependent pathway. J Leukoc Biol 83:1484–1492
Grigorian M et al (2001) Tumor suppressor p53 protein is a new target for the metastasis-associated Mts1/S100A4 protein: functional consequences of their interaction. J Biol Chem 276:22699–22708
Helfman DM, Kim EJ, Lukanidin E, Grigorian M (2005) The metastasis associated protein S100A4: role in tumour progression and metastasis. Br J Cancer 92:1955–1958
Hofmann MA et al (1999) RAGE mediates a novel proinflammatory axis: a central cell surface receptor for S100/calgranulin polypeptides. Cell 97:889–901
Hoppmann S, Steinbach J, Pietzsch J (2010) Scavenger receptors are associated with cellular interactions of S100A12 in vitro and in vivo. Int J Biochem Cell Biol 42:651–661
Hoppmann S, Haase C, Richter S, Pietzsch J (2008a) Expression, purification and fluorine-18 radiolabeling of recombinant S100 proteins—potential probes for molecular imaging of receptor for advanced glycation endproducts (RAGE) in vivo. Protein Expr Purif 57:143–152
Hoppmann S, Haase C, Richter S, Strobel K, Steinbach J, Pietzsch J (2008b) Fluorine-18 radiolabeling of S100/calgranulins: potential probes for molecular imaging of receptor for advanced glycation endproducts (RAGE) in vivo. In: Chen X (ed) Recent advances of bioconjugation chemistry in molecular imaging. Research Signpost, Trivandrum, pp 329–351
Hsieh HL, Schafer BW, Weigle B, Heizmann CW (2004) S100 protein translocation in response to extracellular S100 is mediated by receptor for advanced glycation endproducts in human endothelial cells. Biochem Biophys Res Commun 316:949–959
Huttunen HJ, Kuja-Panula J, Sorci G, Agneletti AL, Donato R, Rauvala H (2000) Coregulation of neurite outgrowth and cell survival by amphoterin and S100 proteins through receptor for advanced glycation end products (RAGE) activation. J Biol Chem 275:40096–40105
Kiryushko D, Novitskaya V, Soroka V, Klingelhofer J, Lukanidin E, Berezin V, Bock E (2006) Molecular mechanisms of Ca(2+) signaling in neurons induced by the S100A4 protein. Mol Cell Biol 26:3625–3638
Leclerc E, Fritz G, Weibel M, Heizmann CW, Galichet A (2007) S100B and S100A6 differentially modulate cell survival by interacting with distinct RAGE (receptor for advanced glycation end products) immunoglobulin domains. J Biol Chem 282:31317–31331
Leclerc E, Heizmann CW, Vetter SW (2009a) RAGE and S100 protein transcription levels are highly variable in human melanoma tumors and cells. Gen Physiol Biophys 28(Spec No Focus):F65–F75
Leclerc E, Fritz G, Vetter SW, Heizmann CW (2009b) Binding of S100 proteins to RAGE: an update. Biochim Biophys Acta 1793(6):993–1007
Li ZH, Dulyaninova NG, House RP, Almo SC, Bresnick AR (2010) S100A4 regulates macrophage chemotaxis. Mol Biol Cell 21:2598–2610
Logsdon CD, Fuentes MK, Huang EH, Arumugam T (2007) RAGE and RAGE ligands in cancer. Curr Mol Med 7:777–789
Miranda KJ, Loeser RF, Yammani RR (2010) Sumoylation and nuclear translocation of S100A4 regulate IL-1beta-mediated production of matrix metalloproteinase-13. J Biol Chem 285(41):31517–31524
Pietzsch J, Bergmann R, Wuest F, Pawelke B, Hultsch C, van den Hoff J (2005) Catabolism of native and oxidized low density lipoproteins: in vivo insights from small animal positron emission tomography studies. Amino Acids 29:389–404
Robinson MJ, Tessier P, Poulsom R, Hogg N (2002) The S100 family heterodimer, MRP-8/14, binds with high affinity to heparin and heparan sulfate glycosaminoglycans on endothelial cells. J Biol Chem 277:3658–3665
Salama I, Malone PS, Mihaimeed F, Jones JL (2008) A review of the S100 proteins in cancer. Eur J Surg Oncol 34(4):357–364
Schmidt AM, Yan SD, Wautier JL, Stern D (1999) Activation of receptor for advanced glycation end products: a mechanism for chronic vascular dysfunction in diabetic vasculopathy and atherosclerosis. Circ Res 84:489–497
Semov A, Moreno MJ, Onichtchenko A, Abulrob A, Ball M, Ekiel I, Pietrzynski G, Stanimirovic D, Alakhov V (2005) Metastasis-associated protein S100A4 induces angiogenesis through interaction with Annexin II and accelerated plasmin formation. J Biol Chem 280:20833–20841
Sherbet GV (2009) Metastasis promoter S100A4 is a potentially valuable molecular target for cancer therapy. Cancer Lett 280:15–30
Stern D, Yan SD, Yan SF, Schmidt AM (2002) Receptor for advanced glycation endproducts: a multiligand receptor magnifying cell stress in diverse pathologic settings. Adv Drug Deliv Rev 54:1615–1625
Tarabykina S, Griffiths TR, Tulchinsky E, Mellon JK, Bronstein IB, Kriajevska M (2007) Metastasis-associated protein S100A4: spotlight on its role in cell migration. Curr Cancer Drug Targets 7:217–228
Wolf R et al (2008) Chemotactic activity of S100A7 (Psoriasin) is mediated by the receptor for advanced glycation end products and potentiates inflammation with highly homologous but functionally distinct S100A15. J Immunol 181:1499–1506
Wolfe RR (1992) Radioactive and stable isotope tracers in biomedicine: principles and practice of kinetic analysis. Wiley Liss, New York
Yammani RR, Carlson CS, Bresnick AR, Loeser RF (2006) Increase in production of matrix metalloproteinase 13 by human articular chondrocytes due to stimulation with S100A4: role of the receptor for advanced glycation end products. Arthritis Rheum 54:2901–2911
Zimmer DB, Wright Sadosky P, Weber DJ (2003) Molecular mechanisms of S100-target protein interactions. Microsc Res Tech 60:552–559
Acknowledgments
The authors are grateful to Mareike Barth, Catharina Heinig, Regina Herrlich, Uta Lenkeit, and Aline Morgenegg for their expert technical assistance. The authors thank Torsten Kniess, Ph.D., and Birgit Mosch, Ph.D. for their expert advice and many stimulating discussions. This work in part was supported by a grant from the Deutsche Forschungsgemeinschaft (DFG; grant no. PI304/1-1).
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Wolf, S., Haase-Kohn, C., Lenk, J. et al. Expression, purification and fluorine-18 radiolabeling of recombinant S100A4: a potential probe for molecular imaging of receptor for advanced glycation endproducts in vivo?. Amino Acids 41, 809–820 (2011). https://doi.org/10.1007/s00726-010-0822-x
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DOI: https://doi.org/10.1007/s00726-010-0822-x