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PDBsum entry 3fj8

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protein ligands Protein-protein interface(s) links
Hormone PDB id
3fj8
Jmol
Contents
Protein chains
141 a.a. *
Ligands
SO4 ×3
Waters ×187
* Residue conservation analysis
PDB id:
3fj8
Name: Hormone
Title: Crystal structure of c117i mutant of human acidic fibroblast growth factor
Structure: Heparin-binding growth factor 1. Chain: a, b. Synonym: hbgf-1, acidic fibroblast growth factor, afgf, beta-endothelial cell growth factor, ecgf-beta. Engineered: yes. Mutation: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Gene: fgf1, fgfa. Expressed in: escherichia coli. Expression_system_taxid: 562.
Resolution:
2.00Å     R-factor:   0.198     R-free:   0.231
Authors: M.Blaber,J.Lee
Key ref:
J.Lee and M.Blaber (2009). The interaction between thermodynamic stability and buried free cysteines in regulating the functional half-life of fibroblast growth factor-1. J Mol Biol, 393, 113-127. PubMed id: 19695265 DOI: 10.1016/j.jmb.2009.08.026
Date:
14-Dec-08     Release date:   06-Oct-09    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chains
Pfam   ArchSchema ?
P05230  (FGF1_HUMAN) -  Fibroblast growth factor 1
Seq:
Struc:
155 a.a.
141 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 5 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     extracellular region   8 terms 
  Biological process     Fc-epsilon receptor signaling pathway   26 terms 
  Biochemical function     S100 protein binding     7 terms  

 

 
DOI no: 10.1016/j.jmb.2009.08.026 J Mol Biol 393:113-127 (2009)
PubMed id: 19695265  
 
 
The interaction between thermodynamic stability and buried free cysteines in regulating the functional half-life of fibroblast growth factor-1.
J.Lee, M.Blaber.
 
  ABSTRACT  
 
Protein biopharmaceuticals are an important and growing area of human therapeutics; however, the intrinsic property of proteins to adopt alternative conformations (such as during protein unfolding and aggregation) presents numerous challenges, limiting their effective application as biopharmaceuticals. Using fibroblast growth factor-1 as model system, we describe a cooperative interaction between the intrinsic property of thermostability and the reactivity of buried free-cysteine residues that can substantially modulate protein functional half-life. A mutational strategy that combines elimination of buried free cysteines and secondary mutations that enhance thermostability to achieve a substantial gain in functional half-life is described. Furthermore, the implementation of this design strategy utilizing stabilizing mutations within the core region resulted in a mutant protein that is essentially indistinguishable from wild type as regard protein surface and solvent structure, thus minimizing the immunogenic potential of the mutations. This design strategy should be generally applicable to soluble globular proteins containing buried free-cysteine residues.
 
  Selected figure(s)  
 
Figure 2.
Fig. 2. Relaxed stereo diagrams of the Leu44 → Trp mutant (a), Phe85 → Trp mutant (b), Phe132 → Trp mutant (c), Val31 → Ile mutant (d), and Cys117 → Ile mutant (e) overlaid onto the wild-type FGF-1 (PDB code 1JQZ) structure (dark gray). Also shown are cavities adjacent to these mutant positions within the wild-type structure that are detectable using a 1.2-Å-radius probe (see Fig. 1 for details).
Figure 3.
Fig. 3. Relaxed stereo diagrams of the Leu44 → Phe/Phe132 → Trp double mutant (a) and the Leu44 → Phe/Cys83 → Thr/Cys117 → Val/Phe132 → Trp quadruple mutant (b) overlaid onto the wild-type (PDB code 1JQZ) FGF-1 structure (dark gray). (a) also shows the location of the cavities (cav4, cav5, and cav6) that are either filled, or partially filled, in response to the Leu44 → Phe/Phe132 → Trp double mutation (partially-filled cav2 is omitted for clarity). In (b), the overlay with the wild-type structure shows only the main-chain atoms within 5 Å of positions 44, 83, 117, and 132.
 
  The above figures are reprinted by permission from Elsevier: J Mol Biol (2009, 393, 113-127) copyright 2009.  
  Figures were selected by an automated process.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
21315087 J.Lee, S.I.Blaber, V.K.Dubey, and M.Blaber (2011).
A polypeptide "building block" for the β-trefoil fold identified by "top-down symmetric deconstruction".
  J Mol Biol, 407, 744-763.
PDB code: 3o3q
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