title: Control of fibroblast growth factor (FGF) 7- and FGF1-induced mitogenesis and downstream signaling by distinct heparin octasaccharide motifs.
abstract: Variation in length, disaccharide composition, and sulfation of heparan sulfate (HS) affects fibroblast growth factor (FGF) signaling. However, it is unclear whether the specific distribution of groups within oligosaccharides or random variations in charge density underlies the effects. Recently we showed that a mixture of undersulfated octasaccharides exhibiting 7 and 8 sulfates (7,8-S-OctaF7) generated from heparin had the highest affinity for FGF7 monitored by salt resistance (>0.60 M salt) of octasaccharide-FGF7 complexes. 7,8-S-OctaF7 also had the highest specific activity for formation of a complex with dimeric FGFR2IIIb competent to bind FGF7. Here we show that when endogenous HS was inhibited by chlorate treatment, 7,8-S-OctaF7 specifically supported FGF7-stimulated DNA synthesis and downstream signaling in FGFR2IIIb-expressing mouse keratinocytes. It failed to support FGF1 signaling in both HS-deficient mouse keratinocytes and 3T3 fibroblasts. In contrast, abundant, more highly sulfated and heterogenous mixtures of octasaccharides with lower affinity (0.30-0.60 M salt) for FGF7 supported FGF1-induced signaling in both cell types. In contrast to the two-component 7,8-S-OctaF7 mixture from FGF7, the high affinity octasaccharide fraction from FGF1 was a heterogeneous mixture with components ranging from 8 to 12 sulfates with 11-S-octasaccharides the most abundant. The high affinity fraction exhibited similar properties to the lower affinity fractions from both FGF1 and FGF7. Octasaccharide mixtures eluting from FGF1 between 0.30 and 0.60 M and above 0.60 M salt were nearly equal in support of FGF1 signaling in fibroblasts and keratinocytes. Both were deficient in support of FGF7-induced signaling in keratinocytes. The results show that both variations in overall charge density and specific distribution of charged groups within HS motifs exhibit FGF-specific control over formation of FGF-HS-FGFR complexes and downstream signaling.
authors.affiliation: Center for Cancer Biology and Nutrition, Institute of Biosciences and Technology, The Texas A&M University System Health Science Center, Houston, Texas 77030-3303.
authors.forename: Yongde
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Department of Physiology, The University of Texas Southwestern Medical Center, Dallas, Texas 75390-9040.
authors.forename: Sheng
authors.initials: S
authors.lastname: Ye
authors.affiliation: Zeria Pharmaceutical Co., Ltd., GS PlatZ., 2512-1, Oshikiri, Kohnan-Machi, Ohsato-Gun, Saitama 360-0111, Japan.
authors.forename: Mikio
authors.initials: M
authors.lastname: Kan
authors.affiliation: Center for Cancer Biology and Nutrition, Institute of Biosciences and Technology, The Texas A&M University System Health Science Center, Houston, Texas 77030-3303; Department of Biochemistry and Biophysics, Texas A&M University, Houston, Texas 77030-3303. Electronic address: wmckeehan@ibt.tamhsc.edu.
authors.forename: Wallace L
authors.initials: WL
authors.lastname: McKeehan
mesh_terms: D016475:3T3 Cells; D020816:Amino Acid Motifs; D000818:Animals; D004247:DNA / Q000737:chemistry; D019281:Dimerization; D016220:Fibroblast Growth Factor 1 / Q000096:biosynthesis* / Q000737:chemistry; D051523:Fibroblast Growth Factor 7 / Q000096:biosynthesis*; D006493:Heparin / Q000737:chemistry*; D015603:Keratinocytes / Q000378:metabolism; D051379:Mice; D009844:Oligosaccharides / Q000737:chemistry*; D051497:Receptor, Fibroblast Growth Factor, Type 2 / Q000737:chemistry; D015398:Signal Transduction
chemical_list: C496454:Fgf7 protein, mouse; D009844:Oligosaccharides; D016220:Fibroblast Growth Factor 1; D051523:Fibroblast Growth Factor 7; D006493:Heparin; D004247:DNA; D051497:Receptor, Fibroblast Growth Factor, Type 2; C065104:keratinocyte growth factor receptor