PDBsum entry 3a6q

Electron transport

PDB id: 3a6q

Contents

Protein chains

122 a.a. *

Ligands

FMN ×2

Metals

_CL ×2

Waters ×396

* Residue conservation analysis

PDB id:

3a6q

Name:

Electron transport

Title:

E13t mutant of fmn-binding protein from desulfovibrio vulgaris (miyazaki f)

Structure:

Fmn-binding protein. Chain: a, b. Engineered: yes. Mutation: yes

Source:

Desulfovibrio vulgaris. Organism_taxid: 883. Strain: miyazaki f. Gene: dvmf_2023. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.40Å R-factor: 0.157 R-free: 0.182

Authors:

T.Nakanishi,Y.Haruyama,H.Inoue,M.Kitamura

Key ref:

H.Chosrowjan et al. (2010). Effects of the disappearance of one charge on ultrafast fluorescence dynamics of the FMN binding protein. J Phys Chem B, 114, 6175-6182. PubMed id: 20397678

Date:

08-Sep-09 Release date: 01-Sep-10

Protein chains

Q46604  (FMNB_DESVM) -  FMN-binding protein from Desulfovibrio vulgaris (strain DSM 19637 / Miyazaki F)

Seq: 122 a.a.

Struc: 122 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

J Phys Chem B 114:6175-6182 (2010)

PubMed id: 20397678

Effects of the disappearance of one charge on ultrafast fluorescence dynamics of the FMN binding protein.

H.Chosrowjan, S.Taniguchi, N.Mataga, T.Nakanishi, Y.Haruyama, S.Sato, M.Kitamura, F.Tanaka.

ABSTRACT

Crystal structures of E13T (Glu13 was replaced by Thr13) and E13Q (Glu13 was replaced by Gln13) FMN binding proteins (FMN-bp) from Desulfovibrio vulgaris, strain Miyazaki F, were determined by the X-ray diffraction method. Geometrical factors related to photoinduced electron transfer from Trp32, Tyr35, and Trp106 to the excited isoalloxazine (Iso*) were compared among the three forms of FMN-bp. The rate of ET is considered to be fastest from Trp32 to Iso* in FMN-bp and then from Tyr35 and Trp106. The distances between Iso and Trp32 did not change appreciably (0.705-0.712 nm) among WT, E13T, and E13Q FMN-bps, though the distances between Iso and Tyr35 or Trp106 became a little shorter by ca. 0.01 nm in both mutated FMN-bps. The distances between the residue at 13 and the ET donors or acceptor in the mutated proteins, however, changed markedly, compared to WT. Hydrogen bonding pairs and distances between Iso and surrounding amino acids were not modified when Glu13 was replaced by Thr13 or Gln13. Effects of elimination of ionic charge at Glu13 on the ultrafast fluorescence dynamics in E13T and E13Q were investigated comparing to WT, by means of a fluorescence up-conversion method. Fluorescence lifetimes were tau(1) = 107 fs (alpha(1) = 0.86), tau(2) = 475 fs (alpha(2) = 0.12), and tau(3) = 30 ps (alpha(3) = 0.02) in E13T and tau(1) = 134 fs (alpha(1) = 0.85), alpha(2) = 746 fs (alpha(2) = 0.12), and tau(3) = 30 ps (alpha(3) = 0.03) in E13Q, which are compared to the reported lifetimes in WT, tau(1) = 168 fs (alpha(1) = 0.95) and alpha(2) = 1.4 ps (alpha(2) = 0.05). Average lifetimes (tau(AV) = Sigma(i=1)(2or3)alpha(i)tau(i)) were 0.75 ps in E13T, 1.10 ps in E13Q, and 0.23 ps in WT, which implies that tau(AV) was 3.3 times longer in E13T and 4.8 times longer in E13Q, compared to WT. The ultrafast fluorescence dynamics of WT did not change when solvent changed from H(2)O to D(2)O. Static ET rates (inverse of average lifetimes) were analyzed with static structures of the three systems of FMN-bp. Net electrostatic (ES) energies of Iso and Trp32, on which ET rates depend, were 0.0263 eV in WT, 0.322 eV in E13T, and 0.412 eV in E13Q. The calculated ET rates were in excellent agreement with the observed ones in all systems.