Standard

Distant non-obvious mutations influence the activity of a hyperthermophilic Pyrococcus furiosus phosphoglucose isomerase. / Subramanian, Kalyanasundaram; Mitusińska, Karolina; Raedts, John; Almourfi, Feras; Joosten, Henk Jan; Hendriks, Sjon; Sedelnikova, Svetlana E.; Kengen, Servé W.M.; Hagen, Wilfred R.; More Authors.

In: Biomolecules, Vol. 9, No. 6, 212, 2019.

Research output: Contribution to journalArticleScientificpeer-review

Harvard

Subramanian, K, Mitusińska, K, Raedts, J, Almourfi, F, Joosten, HJ, Hendriks, S, Sedelnikova, SE, Kengen, SWM, Hagen, WR & More Authors 2019, 'Distant non-obvious mutations influence the activity of a hyperthermophilic Pyrococcus furiosus phosphoglucose isomerase' Biomolecules, vol. 9, no. 6, 212. https://doi.org/10.3390/biom9060212

APA

Subramanian, K., Mitusińska, K., Raedts, J., Almourfi, F., Joosten, H. J., Hendriks, S., ... More Authors (2019). Distant non-obvious mutations influence the activity of a hyperthermophilic Pyrococcus furiosus phosphoglucose isomerase. Biomolecules, 9(6), [212]. https://doi.org/10.3390/biom9060212

Vancouver

Subramanian K, Mitusińska K, Raedts J, Almourfi F, Joosten HJ, Hendriks S et al. Distant non-obvious mutations influence the activity of a hyperthermophilic Pyrococcus furiosus phosphoglucose isomerase. Biomolecules. 2019;9(6). 212. https://doi.org/10.3390/biom9060212

Author

Subramanian, Kalyanasundaram ; Mitusińska, Karolina ; Raedts, John ; Almourfi, Feras ; Joosten, Henk Jan ; Hendriks, Sjon ; Sedelnikova, Svetlana E. ; Kengen, Servé W.M. ; Hagen, Wilfred R. ; More Authors. / Distant non-obvious mutations influence the activity of a hyperthermophilic Pyrococcus furiosus phosphoglucose isomerase. In: Biomolecules. 2019 ; Vol. 9, No. 6.

BibTeX

@article{905c7c21d77248cb878c901027d910de,
title = "Distant non-obvious mutations influence the activity of a hyperthermophilic Pyrococcus furiosus phosphoglucose isomerase",
abstract = "The cupin-type phosphoglucose isomerase (PfPGI) from the hyperthermophilic archaeon Pyrococcus furiosus catalyzes the reversible isomerization of glucose-6-phosphate to fructose-6-phosphate. We investigated PfPGI using protein-engineering bioinformatics tools to select functionally-important residues based on correlated mutation analyses. A pair of amino acids in the periphery of PfPGI was found to be the dominant co-evolving mutation. The position of these selected residues was found to be non-obvious to conventional protein engineering methods. We designed a small smart library of variants by substituting the co-evolved pair and screened their biochemical activity, which revealed their functional relevance. Four mutants were further selected from the library for purification, measurement of their specific activity, crystal structure determination, and metal cofactor coordination analysis. Though the mutant structures and metal cofactor coordination were strikingly similar, variations in their activity correlated with their fine-tuned dynamics and solvent access regulation. Alternative, small smart libraries for enzyme optimization are suggested by our approach, which is able to identify non-obvious yet beneficial mutations.",
keywords = "Comulator, Cupin phosphoglucose isomerase, Protein engineering, Pyrococcus furiosus, Solvent access",
author = "Kalyanasundaram Subramanian and Karolina Mitusińska and John Raedts and Feras Almourfi and Joosten, {Henk Jan} and Sjon Hendriks and Sedelnikova, {Svetlana E.} and Kengen, {Serv{\'e} W.M.} and Hagen, {Wilfred R.} and {More Authors}",
year = "2019",
doi = "10.3390/biom9060212",
language = "English",
volume = "9",
journal = "Biomolecules",
issn = "2218-273X",
number = "6",

}

RIS

TY - JOUR

T1 - Distant non-obvious mutations influence the activity of a hyperthermophilic Pyrococcus furiosus phosphoglucose isomerase

AU - Subramanian, Kalyanasundaram

AU - Mitusińska, Karolina

AU - Raedts, John

AU - Almourfi, Feras

AU - Joosten, Henk Jan

AU - Hendriks, Sjon

AU - Sedelnikova, Svetlana E.

AU - Kengen, Servé W.M.

AU - Hagen, Wilfred R.

AU - More Authors, null

PY - 2019

Y1 - 2019

N2 - The cupin-type phosphoglucose isomerase (PfPGI) from the hyperthermophilic archaeon Pyrococcus furiosus catalyzes the reversible isomerization of glucose-6-phosphate to fructose-6-phosphate. We investigated PfPGI using protein-engineering bioinformatics tools to select functionally-important residues based on correlated mutation analyses. A pair of amino acids in the periphery of PfPGI was found to be the dominant co-evolving mutation. The position of these selected residues was found to be non-obvious to conventional protein engineering methods. We designed a small smart library of variants by substituting the co-evolved pair and screened their biochemical activity, which revealed their functional relevance. Four mutants were further selected from the library for purification, measurement of their specific activity, crystal structure determination, and metal cofactor coordination analysis. Though the mutant structures and metal cofactor coordination were strikingly similar, variations in their activity correlated with their fine-tuned dynamics and solvent access regulation. Alternative, small smart libraries for enzyme optimization are suggested by our approach, which is able to identify non-obvious yet beneficial mutations.

AB - The cupin-type phosphoglucose isomerase (PfPGI) from the hyperthermophilic archaeon Pyrococcus furiosus catalyzes the reversible isomerization of glucose-6-phosphate to fructose-6-phosphate. We investigated PfPGI using protein-engineering bioinformatics tools to select functionally-important residues based on correlated mutation analyses. A pair of amino acids in the periphery of PfPGI was found to be the dominant co-evolving mutation. The position of these selected residues was found to be non-obvious to conventional protein engineering methods. We designed a small smart library of variants by substituting the co-evolved pair and screened their biochemical activity, which revealed their functional relevance. Four mutants were further selected from the library for purification, measurement of their specific activity, crystal structure determination, and metal cofactor coordination analysis. Though the mutant structures and metal cofactor coordination were strikingly similar, variations in their activity correlated with their fine-tuned dynamics and solvent access regulation. Alternative, small smart libraries for enzyme optimization are suggested by our approach, which is able to identify non-obvious yet beneficial mutations.

KW - Comulator

KW - Cupin phosphoglucose isomerase

KW - Protein engineering

KW - Pyrococcus furiosus

KW - Solvent access

UR - http://www.scopus.com/inward/record.url?scp=85067175847&partnerID=8YFLogxK

U2 - 10.3390/biom9060212

DO - 10.3390/biom9060212

M3 - Article

VL - 9

JO - Biomolecules

T2 - Biomolecules

JF - Biomolecules

SN - 2218-273X

IS - 6

M1 - 212

ER -

ID: 55116966