Two pyrenylalanines in dihydrofolate reductase form an excimer enabling the study of protein dynamics

Shengxi Chen, Lin Wang, Nour Eddine Fahmi, Stephen J. Benkovic, Sidney Hecht

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Because of the lack of sensitivity to small changes in distance by available FRET pairs (a constraint imposed by the dimensions of the enzyme), a DHFR containing two pyrene moieties was prepared to enable the observation of excimer formation. Pyren-1-ylalanine was introduced into DHFR positions 16 and 49 using an in vitro expression system in the presence of pyren-1-ylalanyl- tRNACUA. Excimer formation (λex 342 nm; λem 481 nm) was observed in the modified DHFR, which retained its catalytic competence and was studied under multiple and single turnover conditions. The excimer appeared to follow a protein conformational change after the H transfer involving the relative position and orientation of the pyrene moieties and is likely associated with product dissociation.

Original languageEnglish (US)
Pages (from-to)18883-18885
Number of pages3
JournalJournal of the American Chemical Society
Volume134
Issue number46
DOIs
StatePublished - Nov 21 2012

Fingerprint

Tetrahydrofolate Dehydrogenase
Pyrene
Proteins
Mental Competency
Enzymes
Observation
pyrene
Oxidoreductases
In Vitro Techniques

ASJC Scopus subject areas

  • Chemistry(all)
  • Catalysis
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Two pyrenylalanines in dihydrofolate reductase form an excimer enabling the study of protein dynamics. / Chen, Shengxi; Wang, Lin; Fahmi, Nour Eddine; Benkovic, Stephen J.; Hecht, Sidney.

In: Journal of the American Chemical Society, Vol. 134, No. 46, 21.11.2012, p. 18883-18885.

Research output: Contribution to journalArticle

@article{24de7041a2d94ff1a0b132cd06295562,
title = "Two pyrenylalanines in dihydrofolate reductase form an excimer enabling the study of protein dynamics",
abstract = "Because of the lack of sensitivity to small changes in distance by available FRET pairs (a constraint imposed by the dimensions of the enzyme), a DHFR containing two pyrene moieties was prepared to enable the observation of excimer formation. Pyren-1-ylalanine was introduced into DHFR positions 16 and 49 using an in vitro expression system in the presence of pyren-1-ylalanyl- tRNACUA. Excimer formation (λex 342 nm; λem 481 nm) was observed in the modified DHFR, which retained its catalytic competence and was studied under multiple and single turnover conditions. The excimer appeared to follow a protein conformational change after the H transfer involving the relative position and orientation of the pyrene moieties and is likely associated with product dissociation.",
author = "Shengxi Chen and Lin Wang and Fahmi, {Nour Eddine} and Benkovic, {Stephen J.} and Sidney Hecht",
year = "2012",
month = "11",
day = "21",
doi = "10.1021/ja307179q",
language = "English (US)",
volume = "134",
pages = "18883--18885",
journal = "Journal of the American Chemical Society",
issn = "0002-7863",
publisher = "American Chemical Society",
number = "46",

}

TY - JOUR

T1 - Two pyrenylalanines in dihydrofolate reductase form an excimer enabling the study of protein dynamics

AU - Chen, Shengxi

AU - Wang, Lin

AU - Fahmi, Nour Eddine

AU - Benkovic, Stephen J.

AU - Hecht, Sidney

PY - 2012/11/21

Y1 - 2012/11/21

N2 - Because of the lack of sensitivity to small changes in distance by available FRET pairs (a constraint imposed by the dimensions of the enzyme), a DHFR containing two pyrene moieties was prepared to enable the observation of excimer formation. Pyren-1-ylalanine was introduced into DHFR positions 16 and 49 using an in vitro expression system in the presence of pyren-1-ylalanyl- tRNACUA. Excimer formation (λex 342 nm; λem 481 nm) was observed in the modified DHFR, which retained its catalytic competence and was studied under multiple and single turnover conditions. The excimer appeared to follow a protein conformational change after the H transfer involving the relative position and orientation of the pyrene moieties and is likely associated with product dissociation.

AB - Because of the lack of sensitivity to small changes in distance by available FRET pairs (a constraint imposed by the dimensions of the enzyme), a DHFR containing two pyrene moieties was prepared to enable the observation of excimer formation. Pyren-1-ylalanine was introduced into DHFR positions 16 and 49 using an in vitro expression system in the presence of pyren-1-ylalanyl- tRNACUA. Excimer formation (λex 342 nm; λem 481 nm) was observed in the modified DHFR, which retained its catalytic competence and was studied under multiple and single turnover conditions. The excimer appeared to follow a protein conformational change after the H transfer involving the relative position and orientation of the pyrene moieties and is likely associated with product dissociation.

UR - http://www.scopus.com/inward/record.url?scp=84869384366&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84869384366&partnerID=8YFLogxK

U2 - 10.1021/ja307179q

DO - 10.1021/ja307179q

M3 - Article

VL - 134

SP - 18883

EP - 18885

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

SN - 0002-7863

IS - 46

ER -