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PDBsum entry 6f1f
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Blood clotting
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PDB id
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6f1f
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PDB id:
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| Name: |
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Blood clotting
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Title:
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The methylene thioacetal bpti (bovine pancreatic trypsin inhibitor) mutant structure
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Structure:
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Pancreatic trypsin inhibitor. Chain: a, b, c, d, e. Synonym: aprotinin,basic protease inhibitor,bpti. Engineered: yes
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Source:
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Synthetic: yes. Bos taurus. Bovine. Organism_taxid: 9913
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Resolution:
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1.72Å
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R-factor:
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0.190
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R-free:
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0.207
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Authors:
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S.Lansky,R.Mousa,N.Metanis,G.Shoham
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Key ref:
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R.Mousa
et al.
(2018).
BPTI folding revisited: switching a disulfide into methylene thioacetal reveals a previously hidden path.
Chem Sci,
9,
4814-4820.
PubMed id:
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Date:
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21-Nov-17
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Release date:
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18-Jul-18
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PROCHECK
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Headers
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References
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P00974
(BPT1_BOVIN) -
Pancreatic trypsin inhibitor from Bos taurus
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Seq:
Struc:
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100 a.a.
57 a.a.*
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Key: |
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Secondary structure |
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*
PDB and UniProt seqs differ
at 1 residue position (black
cross)
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Chem Sci
9:4814-4820
(2018)
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PubMed id:
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BPTI folding revisited: switching a disulfide into methylene thioacetal reveals a previously hidden path.
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R.Mousa,
S.Lansky,
G.Shoham,
N.Metanis.
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ABSTRACT
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Bovine pancreatic trypsin inhibitor (BPTI) is a 58-residue protein that is
stabilized by three disulfide bonds at positions 5-55, 14-38 and 30-51. Widely
studied for about 50 years, BPTI represents a folding model for many
disulfide-rich proteins. In the study described below, we replaced the solvent
exposed 14-38 disulfide bond with a methylene thioacetal bridge in an attempt to
arrest the folding pathway of the protein at its two well-known intermediates,
N' and N*. The modified protein was expected to be unable to undergo the
rate-determining step in the widely accepted BPTI folding mechanism: the opening
of the 14-38 disulfide bond followed by rearrangements that leads to the native
state, N. Surprisingly, instead of halting BPTI folding at N' and N*, we
uncovered a hidden pathway involving a direct reaction between the N*
intermediate and the oxidizing reagent glutathione (GSSG) to form the
disulfide-mixed intermediate N*-SG, which spontaneously folds into N. On the
other hand, N' was unable to fold into N. In addition, we found that the
methylene thioacetal bridge enhances BPTI stability while fully maintaining its
structure and biological function. These findings suggest a general strategy for
enhancing protein stability without compromising on function or structure,
suggesting potential applications for future therapeutic protein production.
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Links
