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PDBsum entry 2i9v

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Signaling protein PDB id
2i9v
Jmol
Contents
Protein chain
125 a.a. *
Ligands
HC4
Waters ×70
* Residue conservation analysis
PDB id:
2i9v
Name: Signaling protein
Title: Structural role of y98 in pyp: effects on fluorescence, gateway and photocycle recovery
Structure: Photoactive yellow protein. Chain: a. Synonym: pyp. Engineered: yes. Mutation: yes
Source: Halorhodospira halophila. Organism_taxid: 1053. Gene: pyp. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.
Resolution:
2.20Å     R-factor:   0.199     R-free:   0.241
Authors: S.N.Savvides,J.Kyndt
Key ref:
J.A.Kyndt et al. (2007). Structural role of tyrosine 98 in photoactive yellow protein: effects on fluorescence, gateway, and photocycle recovery. Biochemistry, 46, 95. PubMed id: 17198379 DOI: 10.1021/bi061867y
Date:
06-Sep-06     Release date:   06-Feb-07    
PROCHECK
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 Headers
 References

Protein chain
Pfam   ArchSchema ?
P16113  (PYP_HALHA) -  Photoactive yellow protein
Seq:
Struc:
125 a.a.
125 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     response to stimulus   5 terms 
  Biochemical function     signal transducer activity     2 terms  

 

 
DOI no: 10.1021/bi061867y Biochemistry 46:95 (2007)
PubMed id: 17198379  
 
 
Structural role of tyrosine 98 in photoactive yellow protein: effects on fluorescence, gateway, and photocycle recovery.
J.A.Kyndt, S.N.Savvides, S.Memmi, M.Koh, J.C.Fitch, T.E.Meyer, M.P.Heyn, J.J.Van Beeumen, M.A.Cusanovich.
 
  ABSTRACT  
 
We have recently shown that the Y98Q mutant of PYP has a major effect on the photocycle kinetics ( approximately 40 times slower recovery). We have now determined the crystal structure of Y98Q at 2.2 A resolution to reveal the role of residue Y98 in the PYP photocycle. Although the overall structure is very similar to that of WT, we observed two major effects of the mutation. One obvious consequence is a conformational change of the beta4-beta5 loop, which includes a repositioning of residue M100. It had previously been shown that the photocycle is slowed by as much as 3 orders of magnitude when residue M100 is substituted or when the conformation is altered as in Rhodocista centenaria PYP. To investigate whether the altered photocycle of Y98Q is due to this repositioning of M100 or is caused by an effect unrelated to M100, we determined the dark recovery kinetics of the Y98Q/M100A mutant. We find the recovery kinetics to be very similar to the M100A single mutant kinetics and therefore conclude that the slower recovery kinetics in Y98Q are most likely due to repositioning of M100. In addition, we find that other substitutions at position 98 (Y98W, Y98L, and Y98A) have differing effects on the photocycle recovery, presumably due to a variable distortion of the beta4-beta5 loop. The second effect of the Y98Q mutation is a repositioning of R52, which is thought to interact with Y98 in WT PYP and now forms new interactions with residues Q99 and Q56. To determine the role of R52, we also characterized an R52A/M100A double mutant and found that the effects on the recovery kinetics ( approximately 2000 slower recovery than WT) are due to unrelated events in the photocycle. Since the Y98Q/M100A recovery kinetics are more similar to those of M100 than R52A/M100A, we conclude that the repositioning of R52, caused by the Y98Q mutation, does not affect the dark state recovery. In addition, it has been proposed that Y98 and P68 are "gateway residues" between which the chromophore must pass during isomerization. We tested the recovery kinetics of mutant P68A and found that, although the gateway may be important for photocycle initiation, its role in recovery to the ground state is minimal.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
  20953237 E.C.Carroll, S.H.Song, M.Kumauchi, I.H.van Stokkum, A.Jailaubekov, W.D.Hoff, and D.S.Larsen (2010).
Subpicosecond Excited-State Proton Transfer Preceding Isomerization During the Photorecovery of Photoactive Yellow Protein.
  J Phys Chem Lett, 1, 2793-2799.  
18399917 M.Kumauchi, M.T.Hara, P.Stalcup, A.Xie, and W.D.Hoff (2008).
Identification of six new photoactive yellow proteins--diversity and structure-function relationships in a bacterial blue light photoreceptor.
  Photochem Photobiol, 84, 956-969.  
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