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PDBsum entry 1a54
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Phosphotransferase PDB id
1a54

 

 

 

 

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Contents
Protein chain
321 a.a. *
Ligands
2HP
MDC
Waters ×334
* Residue conservation analysis
PDB id:
1a54
Name: Phosphotransferase
Title: Phosphate-binding protein mutant a197c labelled with a coumarin fluorophore and bound to dihydrogenphosphate ion
Structure: Phosphate-binding protein psts. Chain: a. Engineered: yes. Mutation: yes
Source: Escherichia coli. Organism_taxid: 562. Cell_line: an2538. Cellular_location: periplasm. Gene: psts, bvcmskkp036_01783, eyx47_15990. Expressed in: escherichia coli. Expression_system_taxid: 562. Expression_system_cell_line: an2538.
Resolution:
1.60Å     R-factor:   0.177     R-free:   0.208
Authors: M.Hirshberg,K.Henrick,L.Lloyd-Haire,N.Vasisht,M.Brune,J.E.T.Corrie, M.R.Webb
Key ref:
M.Hirshberg et al. (1998). Crystal structure of phosphate binding protein labeled with a coumarin fluorophore, a probe for inorganic phosphate. Biochemistry, 37, 10381-10385. PubMed id: 9671506 DOI: 10.1021/bi980428z
Date:
19-Feb-98     Release date:   14-Oct-98    
PROCHECK
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 Headers
 References

Protein chain
P0AG82  (PSTS_ECOLI) -  Phosphate-binding protein PstS from Escherichia coli (strain K12)
Seq:
Struc: 		346 a.a.
321 a.a.*
Key:    Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 

 
DOI no: 10.1021/bi980428z Biochemistry 37:10381-10385 (1998)
PubMed id: 9671506  
 
 
Crystal structure of phosphate binding protein labeled with a coumarin fluorophore, a probe for inorganic phosphate.
M.Hirshberg, K.Henrick, L.L.Haire, N.Vasisht, M.Brune, J.E.Corrie, M.R.Webb.
 
  ABSTRACT  
 
Crystal structures are presented for the A197C mutant of Escherichia coli phosphate binding protein (PBP) and the same mutant labeled at Cys197 with N-[2-(1-maleimidyl)ethyl]-7-(diethylamino)coumarin-3-carboxamide (MDCC). Both proteins are complexed with inorganic phosphate. The latter molecule, MDCC-PBP, exhibits a large increase in fluorescence on binding inorganic phosphate. The resulting high-fluorescence state of the coumarin and the ability of this coumarin to monitor the conformational changes associated with inorganic phosphate binding are interpreted in terms of the specific interactions of MDCC with the protein. The structure helps to explain why this particular label gives a high-fluorescence state on binding inorganic phosphate, while several other related labels do not, and hence aids our general understanding of environmentally sensitive fluorescence probes on proteins.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
19693761 H.Wang, E.Nakata, and I.Hamachi (2009).
Recent progress in strategies for the creation of protein-based fluorescent biosensors.
  Chembiochem, 10, 2560-2577.  
19199818 J.L.Hougland, C.L.Lamphear, S.A.Scott, R.A.Gibbs, and C.A.Fierke (2009).
Context-dependent substrate recognition by protein farnesyltransferase.
  Biochemistry, 48, 1691-1701.  
18834851 Y.C.Tsai, Z.Jin, and K.A.Johnson (2009).
Site-specific labeling of T7 DNA polymerase with a conformationally sensitive fluorophore and its use in detecting single-nucleotide polymorphisms.
  Anal Biochem, 384, 136-144.  
18535149 A.L.Davidson, E.Dassa, C.Orelle, and J.Chen (2008).
Structure, function, and evolution of bacterial ATP-binding cassette systems.
  Microbiol Mol Biol Rev, 72, 317.  
17372653 M.R.Webb (2007).
Development of fluorescent biosensors for probing the function of motor proteins.
  Mol Biosyst, 3, 249-256.  
17089349 D.Altschuh, S.Oncul, and A.P.Demchenko (2006).
Fluorescence sensing of intermolecular interactions and development of direct molecular biosensors.
  J Mol Recognit, 19, 459-477.  
17034793 H.Gu, S.Lalonde, S.Okumoto, L.L.Looger, A.M.Scharff-Poulsen, A.R.Grossman, J.Kossmann, I.Jakobsen, and W.B.Frommer (2006).
A novel analytical method for in vivo phosphate tracking.
  FEBS Lett, 580, 5885-5893.  
15313245 M.A.Dwyer, and H.W.Hellinga (2004).
Periplasmic binding proteins: a versatile superfamily for protein engineering.
  Curr Opin Struct Biol, 14, 495-504.  
12077430 M.D.Purdy, P.Ge, J.Chen, P.R.Selvin, and M.C.Wiener (2002).
Thiol-reactive lanthanide chelates for phasing protein X-ray diffraction data.
  Acta Crystallogr D Biol Crystallogr, 58, 1111-1117.  
12356327 M.J.Blackman, J.E.Corrie, J.C.Croney, G.Kelly, J.F.Eccleston, and D.M.Jameson (2002).
Structural and biochemical characterization of a fluorogenic rhodamine-labeled malarial protease substrate.
  Biochemistry, 41, 12244-12252.  
12381848 R.M.de Lorimier, J.J.Smith, M.A.Dwyer, L.L.Looger, K.M.Sali, C.D.Paavola, S.S.Rizk, S.Sadigov, D.W.Conrad, L.Loew, and H.W.Hellinga (2002).
Construction of a fluorescent biosensor family.
  Protein Sci, 11, 2655-2675.  
12115121 S.Shrestha, L.L.Salins, C.Mark Ensor, S.Daunert, and S.Daunert (2002).
Rationally designed fluorescently labeled sulfate-binding protein mutants: evaluation in the development of a sensing system for sulfate.
  Biotechnol Bioeng, 78, 517-526.  
11509369 M.R.Webb, and J.E.Corrie (2001).
Fluorescent coumarin-labeled nucleotides to measure ADP release from actomyosin.
  Biophys J, 81, 1562-1569.  
11590019 S.J.Russell, F.Gonzalez, L.Joshua-Tor, and S.A.Johnston (2001).
Selective chemical inactivation of AAA proteins reveals distinct functions of proteasomal ATPases.
  Chem Biol, 8, 941-950.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.

 

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