Impact of phenylalanines outside the dimer interface on phosphotriesterase stability and function

Andrew J. Olsen, Leif A. Halvorsen, Ching Yao Yang, Roni Barak Ventura, Liming Yin, P. Douglas Renfrew, Richard Bonneau, Jin Montclare

Research output: Contribution to journalArticle

Abstract

We explore the significance of phenylalanine outside of the phosphotriesterase (PTE) dimer interface through mutagenesis studies and computational modeling. Previous studies have demonstrated that the residue-specific incorporation of para-fluorophenylalanine (pFF) into PTE improves stability, suggesting the importance of phenylalanines in stabilization of the dimer. However, this comes at a cost of decreased solubility due to pFF incorporation into other parts of the protein. Motivated by this, eight single solvent-exposed phenylalanine mutants are evaluated viarosetta and good correspondence between experiments and these predictions is observed. Three residues, F304, F327, and F335, appear to be important for PTE activity and stability, even though they do not reside in the dimer interface region or active site. While the remaining mutants do not significantly affect structure or activity, one variant, F306L, reveals improved activity at ambient and elevated temperatures. These studies provide further insight into role of these residues on PTE function and stability.

Original languageEnglish (US)
Pages (from-to)2092-2106
Number of pages15
JournalMolecular BioSystems
Volume13
Issue number10
DOIs
StatePublished - 2017

Fingerprint

Phosphoric Triester Hydrolases
Phenylalanine
p-Fluorophenylalanine
Mutagenesis
Solubility
Catalytic Domain
Costs and Cost Analysis
Temperature
Proteins

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology

Cite this

Olsen, A. J., Halvorsen, L. A., Yang, C. Y., Barak Ventura, R., Yin, L., Renfrew, P. D., ... Montclare, J. (2017). Impact of phenylalanines outside the dimer interface on phosphotriesterase stability and function. Molecular BioSystems, 13(10), 2092-2106. https://doi.org/10.1039/c7mb00196g

Impact of phenylalanines outside the dimer interface on phosphotriesterase stability and function. / Olsen, Andrew J.; Halvorsen, Leif A.; Yang, Ching Yao; Barak Ventura, Roni; Yin, Liming; Renfrew, P. Douglas; Bonneau, Richard; Montclare, Jin.

In: Molecular BioSystems, Vol. 13, No. 10, 2017, p. 2092-2106.

Research output: Contribution to journalArticle

Olsen AJ, Halvorsen LA, Yang CY, Barak Ventura R, Yin L, Renfrew PD et al. Impact of phenylalanines outside the dimer interface on phosphotriesterase stability and function. Molecular BioSystems. 2017;13(10):2092-2106. https://doi.org/10.1039/c7mb00196g
Olsen, Andrew J. ; Halvorsen, Leif A. ; Yang, Ching Yao ; Barak Ventura, Roni ; Yin, Liming ; Renfrew, P. Douglas ; Bonneau, Richard ; Montclare, Jin. / Impact of phenylalanines outside the dimer interface on phosphotriesterase stability and function. In: Molecular BioSystems. 2017 ; Vol. 13, No. 10. pp. 2092-2106.
@article{6f00a238dd4d4ec6b32263c33f3c4045,
title = "Impact of phenylalanines outside the dimer interface on phosphotriesterase stability and function",
abstract = "We explore the significance of phenylalanine outside of the phosphotriesterase (PTE) dimer interface through mutagenesis studies and computational modeling. Previous studies have demonstrated that the residue-specific incorporation of para-fluorophenylalanine (pFF) into PTE improves stability, suggesting the importance of phenylalanines in stabilization of the dimer. However, this comes at a cost of decreased solubility due to pFF incorporation into other parts of the protein. Motivated by this, eight single solvent-exposed phenylalanine mutants are evaluated viarosetta and good correspondence between experiments and these predictions is observed. Three residues, F304, F327, and F335, appear to be important for PTE activity and stability, even though they do not reside in the dimer interface region or active site. While the remaining mutants do not significantly affect structure or activity, one variant, F306L, reveals improved activity at ambient and elevated temperatures. These studies provide further insight into role of these residues on PTE function and stability.",
author = "Olsen, {Andrew J.} and Halvorsen, {Leif A.} and Yang, {Ching Yao} and {Barak Ventura}, Roni and Liming Yin and Renfrew, {P. Douglas} and Richard Bonneau and Jin Montclare",
year = "2017",
doi = "10.1039/c7mb00196g",
language = "English (US)",
volume = "13",
pages = "2092--2106",
journal = "Molecular BioSystems",
issn = "1742-206X",
publisher = "Royal Society of Chemistry",
number = "10",

}

TY - JOUR

T1 - Impact of phenylalanines outside the dimer interface on phosphotriesterase stability and function

AU - Olsen, Andrew J.

AU - Halvorsen, Leif A.

AU - Yang, Ching Yao

AU - Barak Ventura, Roni

AU - Yin, Liming

AU - Renfrew, P. Douglas

AU - Bonneau, Richard

AU - Montclare, Jin

PY - 2017

Y1 - 2017

N2 - We explore the significance of phenylalanine outside of the phosphotriesterase (PTE) dimer interface through mutagenesis studies and computational modeling. Previous studies have demonstrated that the residue-specific incorporation of para-fluorophenylalanine (pFF) into PTE improves stability, suggesting the importance of phenylalanines in stabilization of the dimer. However, this comes at a cost of decreased solubility due to pFF incorporation into other parts of the protein. Motivated by this, eight single solvent-exposed phenylalanine mutants are evaluated viarosetta and good correspondence between experiments and these predictions is observed. Three residues, F304, F327, and F335, appear to be important for PTE activity and stability, even though they do not reside in the dimer interface region or active site. While the remaining mutants do not significantly affect structure or activity, one variant, F306L, reveals improved activity at ambient and elevated temperatures. These studies provide further insight into role of these residues on PTE function and stability.

AB - We explore the significance of phenylalanine outside of the phosphotriesterase (PTE) dimer interface through mutagenesis studies and computational modeling. Previous studies have demonstrated that the residue-specific incorporation of para-fluorophenylalanine (pFF) into PTE improves stability, suggesting the importance of phenylalanines in stabilization of the dimer. However, this comes at a cost of decreased solubility due to pFF incorporation into other parts of the protein. Motivated by this, eight single solvent-exposed phenylalanine mutants are evaluated viarosetta and good correspondence between experiments and these predictions is observed. Three residues, F304, F327, and F335, appear to be important for PTE activity and stability, even though they do not reside in the dimer interface region or active site. While the remaining mutants do not significantly affect structure or activity, one variant, F306L, reveals improved activity at ambient and elevated temperatures. These studies provide further insight into role of these residues on PTE function and stability.

UR - http://www.scopus.com/inward/record.url?scp=85030104589&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85030104589&partnerID=8YFLogxK

U2 - 10.1039/c7mb00196g

DO - 10.1039/c7mb00196g

M3 - Article

VL - 13

SP - 2092

EP - 2106

JO - Molecular BioSystems

JF - Molecular BioSystems

SN - 1742-206X

IS - 10

ER -