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Engineering protein stability and specificity using fluorous amino acids: the importance of packing effects.

Publication ,  Journal Article
Buer, BC; de la Salud-Bea, R; Al Hashimi, HM; Marsh, ENG
Published in: Biochemistry
November 17, 2009

The incorporation of extensively fluorinated, or fluorous, analogues of hydrophobic amino acids into proteins potentially provides the opportunity to modulate the physicochemical properties of proteins in a predictable manner. On the basis of the properties of small fluorocarbon molecules, extensively fluorinated proteins should be both more thermodynamically stable and self-segregate through "fluorous" interactions between fluorinated amino acids. We have examined the effects of introducing the fluorous leucine analogue l-5,5,5,5',5',5',-hexafluoroleucine (hFLeu) at various positions within the hydrophobic core of a de novo-designed four-alpha-helix bundle protein, alpha(4). The stabilizing effect of hFLeu is strongly dependent on the positions at which it is incorporated, with per-residue DeltaDeltaG(degrees)((fold)) ranging from -0.09 to -0.8 kcal mol(-1) residue(-1). In particular, incorporating hFLeu at all the "a" positions or all the "d" positions of the hydrophobic core, thereby creating an alternating packing arrangement of leucine and hFLeu, leads to very stably folded proteins that exhibit higher per-residue DeltaDeltaG(degrees)((fold)) values than proteins that are packed entirely with hFleu. We conclude that efficient packing of the fluorous amino acid within the hydrophobic core provides a more important contribution to enhancing protein stability than do fluorocarbon-fluorocarbon interactions between fluorinated protein side chains.

Duke Scholars

Published In

Biochemistry

DOI

EISSN

1520-4995

Publication Date

November 17, 2009

Volume

48

Issue

45

Start / End Page

10810 / 10817

Location

United States

Related Subject Headings

  • Thermodynamics
  • Proteins
  • Protein Engineering
  • Nuclear Magnetic Resonance, Biomolecular
  • Models, Molecular
  • Biochemistry & Molecular Biology
  • Amino Acids
  • 3404 Medicinal and biomolecular chemistry
  • 3205 Medical biochemistry and metabolomics
  • 3101 Biochemistry and cell biology
 

Citation

APA
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ICMJE
MLA
NLM
Buer, B. C., de la Salud-Bea, R., Al Hashimi, H. M., & Marsh, E. N. G. (2009). Engineering protein stability and specificity using fluorous amino acids: the importance of packing effects. Biochemistry, 48(45), 10810–10817. https://doi.org/10.1021/bi901481k
Buer, Benjamin C., Roberto de la Salud-Bea, Hashim M. Al Hashimi, and E Neil G. Marsh. “Engineering protein stability and specificity using fluorous amino acids: the importance of packing effects.Biochemistry 48, no. 45 (November 17, 2009): 10810–17. https://doi.org/10.1021/bi901481k.
Buer BC, de la Salud-Bea R, Al Hashimi HM, Marsh ENG. Engineering protein stability and specificity using fluorous amino acids: the importance of packing effects. Biochemistry. 2009 Nov 17;48(45):10810–7.
Buer, Benjamin C., et al. “Engineering protein stability and specificity using fluorous amino acids: the importance of packing effects.Biochemistry, vol. 48, no. 45, Nov. 2009, pp. 10810–17. Pubmed, doi:10.1021/bi901481k.
Buer BC, de la Salud-Bea R, Al Hashimi HM, Marsh ENG. Engineering protein stability and specificity using fluorous amino acids: the importance of packing effects. Biochemistry. 2009 Nov 17;48(45):10810–10817.
Journal cover image

Published In

Biochemistry

DOI

EISSN

1520-4995

Publication Date

November 17, 2009

Volume

48

Issue

45

Start / End Page

10810 / 10817

Location

United States

Related Subject Headings

  • Thermodynamics
  • Proteins
  • Protein Engineering
  • Nuclear Magnetic Resonance, Biomolecular
  • Models, Molecular
  • Biochemistry & Molecular Biology
  • Amino Acids
  • 3404 Medicinal and biomolecular chemistry
  • 3205 Medical biochemistry and metabolomics
  • 3101 Biochemistry and cell biology