Analysis of putative heparin-binding domains of fibroblast growth factor-1. Using site-directed mutagenesis and peptide analogues.


Abstract

The contribution of individual basic amino acids within three putative "consensus sequences" for heparin binding of fibroblast growth factor-1 have been examined by site-directed mutagenesis. The results indicate that a significant reduction in the apparent affinity of fibroblast growth factor-1 for heparin is only observed when basic residues in one of the three regions are mutated. Mutation in the other regions are without affect on heparin binding. The heparin binding properties of synthetic peptides based on the three "consensus sequences" paralleled the mutagenesis results. That is, synthetic peptides corresponding to regions of the protein that were affected by mutagenesis with respect to heparin binding exhibited a relatively high affinity for immobilized heparin, whereas those corresponding to regions of similar charge density that were unaffected by mutagenesis did not. In addition, amino acid substitution of a nonbasic residue in the heparin-binding peptide could abolish its heparin binding capacity. The heparin-binding peptide could antagonize the mitogenic activity of FGF-1, probably because of the heparin dependence of this activity. Together these data demonstrate that the heparin binding properties of fibroblast growth factor-1 are dictated by structural features more complex than clusters of basic amino acids. The results of these and other studies indicate that consensus motifs for heparin-binding require further definition. More importantly, the results provide a basis for the design of peptide-based inhibitors of FGF-1.

Submission Details

ID: 2U6eRS4X3

Submitter: Connie Wang

Submission Date: Sept. 28, 2018, 5:19 p.m.

Version: 1

Publication Details
Wong P;Hampton B;Szylobryt E;Gallagher AM;Jaye M;Burgess WH,J Biol Chem (1995) Analysis of putative heparin-binding domains of fibroblast growth factor-1. Using site-directed mutagenesis and peptide analogues. PMID:7592764
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
1DZC 2000-03-16 High resolution structure of acidic fibroblast growth factor. Mutant FGF-4-ALA-(24-154), 24 NMR structures
2ERM 2006-10-03 Solution structure of a biologically active human FGF-1 monomer, complexed to a hexasaccharide heparin-analogue
1RML 1998-11-11 NMR STUDY OF ACID FIBROBLAST GROWTH FACTOR BOUND TO 1,3,6-NAPHTHALENE TRISULPHONATE, 26 STRUCTURES
1DZD 2000-03-16 High resolution structure of acidic fibroblast growth factor (27-154), 24 NMR structures
2K8R 2009-10-13 Solution structure of human acidic fibroblast growth factor in complex with anti-angiogenic drug inositol hexaphosphate (IP6)
2K4A 2009-06-09 FGF-1-C2A binary complex structure: a key component in the fibroblast growthfactor non-classical pathway
2KI6 2010-03-09 The FGF1-S100A13-C2A hetero-hexameric complex structure: A component in the non-classical pathway for FGF1 secretion
2KI4 2010-03-09 FGF1-S100A13 complex structure: key component in non-classical path way of FGF1
2K43 2009-06-02 Acidic fibroblast growth factor solution structure in the FGF-1-C2A binary complex: key component in the fibroblast growthfactor non-classical pathway
2RQ9 2010-03-16 Solution structure of human acidic fibroblast growth factor (aFGF) in the presence of a protein stabilizer NDSB-new
1RG8 2004-10-05 1.1 Human Acidic Fibroblast Growth Factor (haFGF-1) at 1.10 angstrom resolution (140 amino acid form)
2J3P 2007-02-13 1.4 crystal structure of rat FGF1 at 1.4 A
4QC4 2015-03-11 1.49 Crystal structure of C117S mutant of human acidic fibroblast growth factor
4QBV 2015-03-11 1.5 Crystal structure of C117T mutant of human acidic fibroblast growth factor in sodium citrate buffer
3BB2 2008-04-15 1.5 Crystal structure of L26D/D28N mutant of Human acidic fibroblast growth factor
4QAL 2015-03-11 1.5 Crystal structure of C117A mutant of human acidic fibroblast growth factor
4QBC 2015-03-11 1.52 Crystal structure of C117T mutant of human acidic fibroblast growth factor in sodium formate buffer
3B9U 2008-04-15 1.55 Crystal structure of L26N/D28N/H93G mutant of Human acidic fibroblast growth factor
4Q9G 2015-03-11 1.55 Crystal structure of K12V/C16S/C117V/P134V mutant of human acidic fibroblast growth factor
3BAO 2008-04-15 1.55 Crystal structure of L26N mutant of Human acidic fibroblast growth factor
2HW9 2007-06-12 1.6 Crystal structure of Lys12Cys/Cys117Val mutant of human acidic fibroblast Growth factor at 1.60 angstrom resolution.
3BAU 2008-04-15 1.6 Crystal structure of K12V/L26D/D28A mutant of Human acidic fibroblast growth factor
1P63 2004-05-11 1.6 Human Acidic Fibroblast Growth Factor. 140 Amino Acid Form with Amino Terminal His Tag and Leu111 Replaced with Ile (L111I)
3O3Q 2011-02-23 1.6 Crystal structure of 'L44F/M67I/L73V/A103G/deletion 104-106/F108Y/V109L/L111I/C117V/R119G/deletion 120-122' mutant form of Human acidic fibroblast growth factor
2HWM 2007-06-12 1.6 Crystal structure of Lys12Val/Cys117Val mutant of human acidic fibroblast growth factor at 1.60 angstrom resolution
3BA7 2008-04-15 1.6 Crystal structure of L26N/D28A mutant of Human acidic fibroblast growth factor
3BAV 2008-04-15 1.62 Crystal structure of L26A/D28N mutant of Human acidic fibroblast growth factor
1JQZ 2001-12-19 1.65 Human Acidic Fibroblast Growth Factor. 141 Amino Acid Form with Amino Terminal His Tag.
2HWA 2007-06-12 1.65 Crystal structure of Lys12Thr/Cys117Val mutant of human acidic fibroblast growth factor at 1.65 angstrom resolution.
3BAH 2008-04-15 1.65 Crystal structure of K112N mutant of Human acidic fibroblast growth factor
1M16 2003-08-05 1.7 Human Acidic Fibroblast Growth Factor. 141 Amino Acid Form with Amino Terminal His Tag and Leu 44 Replaced with Phe (L44F), Leu 73 Replaced with Val (L73V), Val 109 Replaced with Leu (V109L) and Cys 117 Replaced with Val (C117V).
1JTC 2001-12-19 1.7 Human Acidic Fibroblast Growth Factor. 141 Amino Acid Form with Amino Terminal His Tag AND LEU 44 REPLACED BY PHE (L44F)
1JY0 2003-08-12 1.7 Human acidic fibroblast growth factor. 141 amino acid form with amino terminal His tag and Cys 117 replaced with Val (C117V).
2HZ9 2007-06-19 1.7 Crystal structure of Lys12Val/Asn95Val/Cys117Val mutant of human acidic fibroblast growth factor at 1.70 angstrom resolution.
2Q9X 2008-07-01 1.7 Crystal structure of highly stable mutant Q40P/S47I/H93G of human fibroblast growth factor-1
1JT7 2001-12-19 1.7 Human Acidic Fibroblast Growth Factor. 141 Amino Acid Form with Amino Terminal His Tag AND LEU 44 REPLACED BY PHE AND LEU 73 REPLACED BY VAL AND VAL 109 REPLACED BY LEU (L44F/L73V/V109L)
3BAG 2008-04-15 1.75 Crystal structure of K112N/N114A mutant of Human acidic fibroblast growth factor
3BA5 2008-04-15 1.75 Crystal structure of D28A mutant of Human acidic fibroblast growth factor
1JT4 2001-12-19 1.78 Human Acidic Fibroblast Growth Factor. 141 Amino Acid Form with Amino Terminal His Tag AND VAL 109 REPLACED BY LEU (V109L)
4Q9P 2015-03-11 1.8 Crystal structure of C16T/K12V/C117V/P134V mutant of human acidic fibroblast growth factor
3BA4 2008-04-15 1.8 Crystal structure of L26D mutant of Human acidic fibroblast growth factor
4Q91 2015-03-11 1.8 Crystal structure of C16A/K12V/C117V/P134V mutant of human acidic fibroblast growth factor
1Q04 2004-07-27 1.8 Crystal structure of FGF-1, S50E/V51N
3BAQ 2008-04-15 1.8 Crystal structure of L26A mutant of Human acidic fibroblast growth factor
3CQA 2009-04-07 1.8 Crystal structure of human fibroblast growth factor-1 with mutations Glu81Ala and Lys101Ala
1JT5 2001-12-19 1.85 Human Acidic Fibroblast Growth Factor. 141 Amino Acid Form with Amino Terminal His Tag AND LEU 73 REPLACED BY VAL AND VAL 109 REPLACED BY LEU (L73V/V109L)
3CRG 2009-02-17 1.85 Crystal structure of human fibroblast growth factor-1 with mutations Glu81Ala, Glu82Asn and Lys101Ala
2AQZ 2006-02-07 1.85 Crystal structure of FGF-1, S17T/N18T/G19 deletion mutant
3FJF 2009-10-06 1.9 Crystal structure of C83T mutant of Human acidic fibroblast growth factor
3FJH 2009-10-06 1.9 Crystal structure of C83A mutant of Human acidic fibroblast growth factor
3FJD 2009-10-06 1.9 Crystal structure of L44F/F132W mutant of Human acidic fibroblast growth factor
1Z2V 2006-02-07 1.9 Crystal Structure of Glu60 deletion Mutant of Human Acidic Fibroblast Growth Factor
3FJJ 2009-10-06 1.9 Crystal structure of C83V mutant of Human acidic fibroblast growth factor
3FJ9 2009-10-06 1.9 Crystal structure of F85W mutant of Human acidic fibroblast growth factor
1NZK 2004-03-02 1.95 Crystal Structure of a Multiple Mutant (L44F, L73V, V109L, L111I, C117V) of Human Acidic Fibroblast Growth Factor
3FGM 2009-10-06 1.95 Crystal structure of L44F/C83T/C117V/F132W mutant of Human acidic fibroblast growth factor
3FJA 2009-10-06 1.95 Crystal structure of F132W mutant of Human acidic fibroblast growth factor
1JT3 2001-12-19 1.95 Human Acidic Fibroblast Growth Factor. 141 Amino Acid Form with Amino Histidine Tag AND LEU 73 REPLACED BY VAL (L73V)
4YOL 2016-03-16 1.97 Human fibroblast growth factor-1 C16S/A66C/C117A/P134A
3K1X 2010-02-02 1.98 Acidic Fibroblast Growth Factor (FGF-1) complexed with dobesilate
1HKN 2004-03-11 2.0 A complex between acidic fibroblast growth factor and 5-amino-2-naphthalenesulfonate
3FJB 2009-10-06 2.0 Crystal structure of V31I mutant of Human acidic fibroblast growth factor
4XKI 2016-01-20 2.0 Human Fibroblast Growth Factor - 1 (FGF-1) mutant S116R
1K5U 2002-02-27 2.0 Human acidic fibroblast growth factor. 141 amino acid form with amino terminal His tag with His93 replaced by Gly (H93G).
2AFG 1995-10-15 2.0 2.0 ANGSTROM X-RAY STRUCTURE OF HUMAN ACIDIC FIBROBLAST GROWTH FACTOR
1PZZ 2004-07-27 2.0 Crystal structure of FGF-1, V51N mutant
3FJ8 2009-10-06 2.0 Crystal structure of C117I mutant of Human acidic fibroblast growth factor
3FJC 2009-10-06 2.0 Crystal structure of L44W mutant of Human acidic fibroblast growth factor
3BAD 2008-04-15 2.0 Crystal structure of D70A/H93G mutant of Human acidic fibroblast growth factor
1Q03 2004-07-27 2.05 Crystal structure of FGF-1, S50G/V51G mutant
4QO3 2015-05-06 2.05 Crystal structure of C16S/N18S/K12V/C117V/P134V mutant of human acidic fibroblast growth factor
3FJE 2009-10-06 2.1 Crystal structure of C83S mutant of Human acidic fibroblast growth factor
1K5V 2002-02-27 2.1 Human acidic fibroblast growth factor. 141 amino acid form with amino terminal His tag with Asn106 replaced by Gly (N106G).
1YTO 2006-01-24 2.1 Crystal Structure of Gly19 deletion Mutant of Human Acidic Fibroblast Growth Factor
3OJM 2011-08-31 2.1 Crystal Structure of FGF1 complexed with the ectodomain of FGFR2b harboring P253R Apert mutation
3CRI 2009-02-17 2.1 Crystal structure of human fibroblast growth factor-1 with mutations Glu81Ser, Glu82Asn and Lys101Ala
3FJK 2009-10-06 2.15 Crystal structure of A66C mutant of Human acidic fibroblast growth factor
3CRH 2009-02-17 2.15 Crystal structure of human fibroblast growth factor-1 with mutations Glu81Ser and Lys101Ala
3OJ2 2011-08-31 2.2 Crystal structure of FGF1 complexed with the ectodomain of FGFR2b harboring the A172F Pfeiffer syndrome mutation
2UUS 2008-05-13 2.2 Crystal structure of the rat FGF1-sucrose octasulfate (SOS) complex.
3JUT 2010-02-02 2.25 Acidic Fibroblast Growth Factor (FGF-1) complexed with gentisic acid
3HOM 2009-10-06 2.3 Crystal structure of oxidized A66C mutant of Human acidic fibroblast growth factor
3UD9 2012-11-28 2.34 Crystal Structure Analysis of FGF1-Disaccharide(NI23) complex
3UD8 2012-11-28 2.37 Crystal Structure Analysis of FGF1-Disaccharide(NI22) complex
1DJS 2000-01-12 2.4 LIGAND-BINDING PORTION OF FIBROBLAST GROWTH FACTOR RECEPTOR 2 IN COMPLEX WITH FGF1
3UDA 2012-11-28 2.51 Crystal Structure Analysis of FGF1-Disaccharide(NI24) complex
2NTD 2007-07-24 2.52 Human fibroblast growth factor-1 (140 amino acid form) with Cys117Val/Pro134Cys mutations
3FJI 2009-10-06 2.55 Crystal structure of K12V/C83I/C117V mutant of Human acidic fibroblast growth factor
1Z4S 2006-02-07 2.6 Crystal Structure of Gly19 and Glu60 deletion mutant of Human Acidic Fibroblast Growth Factor
3OJV 2011-12-28 2.6 Crystal Structure of FGF1 complexed with the ectodomain of FGFR1c exhibiting an ordered ligand specificity-determining betaC'-betaE loop
3CU1 2009-04-21 2.6 Crystal Structure of 2:2:2 FGFR2D2:FGF1:SOS complex
1BAR 1993-10-31 2.7 THREE-DIMENSIONAL STRUCTURES OF ACIDIC AND BASIC FIBROBLAST GROWTH FACTORS
1AFC 1993-10-31 2.7 STRUCTURAL STUDIES OF THE BINDING OF THE ANTI-ULCER DRUG SUCROSE OCTASULFATE TO ACIDIC FIBROBLAST GROWTH FACTOR
1EVT 2000-05-31 2.8 CRYSTAL STRUCTURE OF FGF1 IN COMPLEX WITH THE EXTRACELLULAR LIGAND BINDING DOMAIN OF FGF RECEPTOR 1 (FGFR1)
1E0O 2000-10-23 2.8 CRYSTAL STRUCTURE OF A TERNARY FGF1-FGFR2-HEPARIN COMPLEX
3UD7 2012-11-28 2.8 Crystal Structure Analysis of FGF1-Disaccharide(NI21) complexes
1AXM 1998-04-22 3.0 HEPARIN-LINKED BIOLOGICALLY-ACTIVE DIMER OF FIBROBLAST GROWTH FACTOR
2AXM 1998-04-22 3.0 HEPARIN-LINKED BIOLOGICALLY-ACTIVE DIMER OF FIBROBLAST GROWTH FACTOR
1RY7 2004-02-10 3.2 Crystal Structure of the 3 Ig form of FGFR3c in complex with FGF1
4J23 2014-02-19 3.88 Low resolution crystal structure of the FGFR2D2D3/FGF1/SR128545 complex

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
91.5 Fibroblast growth factor 1 Q9N1S8 FGF1_CAPCA
90.3 Fibroblast growth factor 1 P19596 FGF1_CHICK
90.3 Fibroblast growth factor 1 Q7M303 FGF1_SHEEP
92.3 Fibroblast growth factor 1 P03968 FGF1_BOVIN
95.5 Fibroblast growth factor 1 P61149 FGF1_RAT
95.5 Fibroblast growth factor 1 P61148 FGF1_MOUSE
98.0 Fibroblast growth factor 1 P20002 FGF1_PIG
96.8 Fibroblast growth factor 1 P34004 FGF1_MESAU
100.0 Fibroblast growth factor 1 Q5NVQ3 FGF1_PONAB
100.0 Fibroblast growth factor 1 P05230 FGF1_HUMAN