title: Glycosaminoglycan silencing by engineered CXCL12 variants.
abstract: We have engineered GPCR (G protein-coupled receptor) knock-out and high GAG-binding affinity into CXCL12α to inhibit CXCL12α-induced cell migration. Compared to wtCXCL12, the mutant CXCL12α (Δ8 L29K V39K) exhibited a 5.6-fold and a 2.2-fold affinity increase for heparin and heparan sulfate, respectively. From NaCl-based heparin displacement chromatography we concluded that more amino acid replacements would lead to altered GAG (glycosaminoglycan) ligand specificity. GAG silencing by this mutant was shown in a murine seeding model of human cancer cells, whereby a greatly reduced number of liver metastases was detected when the animals were treated intravenously with 1mg/kg CXCL12α (Δ8 L29K V39K) before cancer cell application.
authors: Institute of Pharmaceutical Sciences, University of Graz, Schubertstraße 1/1, 8010 Graz, Austria.
authors: Martha
authors: M
authors: Gschwandtner
authors: ProtAffin Biotechnologie AG, Graz, Austria.
authors: Martin U
authors: MU
authors: Trinker
authors: Institute of Pharmaceutical Sciences, University of Graz, Schubertstraße 1/1, 8010 Graz, Austria.
authors: Bianca
authors: B
authors: Hecher
authors: ProtAffin Biotechnologie AG, Graz, Austria.
authors: Tiziana
authors: T
authors: Adage
authors: Institute of Cellular Medicine, Newcastle University, Framlington Place, Newcastle upon Tyne NE2 4HH, United Kingdom.
authors: Simi
authors: S
authors: Ali
authors: Institute of Pharmaceutical Sciences, University of Graz, Schubertstraße 1/1, 8010 Graz, Austria; ProtAffin Biotechnologie AG, Graz, Austria. Electronic address: andreas.kungl@uni-graz.at.
authors: Andreas J
authors: AJ
authors: Kungl
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D054377:Chemokine CXCL12 / Q000235:genetics* / Q000378:metabolism; D005260:Female; D020868:Gene Silencing; D006025:Glycosaminoglycans / Q000172:deficiency* / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000473:pathology; D051379:Mice; D009154:Mutation; D015202:Protein Engineering; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics
chemical_list: D054377:Chemokine CXCL12; D006025:Glycosaminoglycans; D043562:Receptors, G-Protein-Coupled
keywords: Cancer therapy; Carbohydrate binding protein; Chemokine; Glycosaminoglycan; Protein engineering; Tumour metastasis