2021
DOI: 10.1073/pnas.2014592118
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Acceleration of catalysis in dihydrofolate reductase by transient, site-specific photothermal excitation

Abstract: We have studied the role of protein dynamics in chemical catalysis in the enzyme dihydrofolate reductase (DHFR), using a pump–probe method that employs pulsed-laser photothermal heating of a gold nanoparticle (AuNP) to directly excite a local region of the protein structure and transient absorbance to probe the effect on enzyme activity. Enzyme activity is accelerated by pulsed-laser excitation when the AuNP is attached close to a network of coupled motions in DHFR (on the FG loop, containing residues 116–132,… Show more

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Cited by 15 publications
(21 citation statements)
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References 57 publications
(52 reference statements)
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“…S1). The majority of the I552A-derived peptides display nearly identical exchange behavior to WT across the 5 temperatures (10,20,25,30, and 40 °C) analyzed (cf. Data S1).…”
Section: Resultsmentioning
confidence: 98%
See 2 more Smart Citations
“…S1). The majority of the I552A-derived peptides display nearly identical exchange behavior to WT across the 5 temperatures (10,20,25,30, and 40 °C) analyzed (cf. Data S1).…”
Section: Resultsmentioning
confidence: 98%
“…S1). The majority of the I552Aderived peptides display nearly identical exchange behavior to WT across the 5 temperatures (10,20,25,30, and 40 °C) analyzed (SI Appendix, Data S1). The peptide of particular interest is 317-334 that represents the surface loop that was previously shown (19) to undergo correlated alterations in the activation energy for HDX Ea(kHDX) and Ea(kcat) when interrogating the variants I553G and L546A in relation to WT.…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…There will be further discussion of this issue later in this Perspective when we discuss laboratory evolved artificial enzymes. While predicted over 11 years ago, this kind of energy transfer from the surface of the protein to the active site has now been observed experimentally in soybean lipoxygenase and also bacterial DHFR for slower motions …”
Section: Rapid Protein Dynamicsmentioning
confidence: 82%
“…Dihydrofolate reductase (DHFR) has been used extensively as a model enzyme for investigating the link between structure, dynamics, and catalysis. The enzyme catalyzes the reduction of 7,8-dihydrofolate (DHF) to 5,6,7,8-tetrahydrofolate (THF) using nicotinamide adenine dinucleotide phosphate (NADPH) as the cofactor (Figure A). Five intermediates have been identified in the steady-state catalytic cycle and characterized by X-ray crystallography and NMR: ,,,, E:NADPH, E:NADPH:DHF, E:NADP + :THF, E:THF, and E:NADPH:THF (Figure B).…”
Section: Introductionmentioning
confidence: 99%