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PDBsum entry 1opd

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Phosphotransferase PDB id
1opd
Jmol
Contents
Protein chain
85 a.a. *
Ligands
SO4
Waters ×54
* Residue conservation analysis
PDB id:
1opd
Name: Phosphotransferase
Title: Histidine-containing protein (hpr), mutant with ser 46 replaced by asp (s46d)
Structure: Histidine-containing protein. Chain: a. Synonym: hpr. Engineered: yes. Mutation: yes
Source: Escherichia coli. Organism_taxid: 562. Strain: esk108. Expressed in: escherichia coli. Expression_system_taxid: 562.
Resolution:
1.50Å     R-factor:   0.182     R-free:   0.234
Authors: S.Napper,L.Delbaere,B.Waygood
Key ref:
S.Napper et al. (1996). Mutation of serine-46 to aspartate in the histidine-containing protein of Escherichia coli mimics the inactivation by phosphorylation of serine-46 in HPrs from gram-positive bacteria. Biochemistry, 35, 11260-11267. PubMed id: 8784179 DOI: 10.1021/bi9603480
Date:
01-Aug-96     Release date:   20-Aug-97    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chain
Pfam   ArchSchema ?
P0AA04  (PTHP_ECOLI) -  Phosphocarrier protein HPr
Seq:
Struc:
85 a.a.
85 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 2 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     cytoplasm   2 terms 
  Biological process     transport   8 terms 
  Biochemical function     protein binding     7 terms  

 

 
DOI no: 10.1021/bi9603480 Biochemistry 35:11260-11267 (1996)
PubMed id: 8784179  
 
 
Mutation of serine-46 to aspartate in the histidine-containing protein of Escherichia coli mimics the inactivation by phosphorylation of serine-46 in HPrs from gram-positive bacteria.
S.Napper, J.W.Anderson, F.Georges, J.W.Quail, L.T.Delbaere, E.B.Waygood.
 
  ABSTRACT  
 
Histidine-containing protein (HPr) is a phosphocarrier protein of the bacterial phosphoenolpyruvate:sugar phosphotransferase system. HPr is phosphorylated at the active site residue, His15, by phosphoenolpyruvate-dependent enzyme I in the first enzyme reaction in the process of phosphoryl transfer to sugar. In many Gram-positive bacterial species HPr may also be phosphorylated at Ser46 by an ATP-dependent protein kinase but not in the Gram-negative Escherichia coli and Salmonella typhimurium. One effect of the phosphorylation at Ser46 is to make HPr a poor acceptor for phosphorylation at His15. In Bacillus subtilis HPr, the mutation Ser46Asp mimics the effects of phosphorylation. A series of mutations were made at Ser46 in E. coli HPr: Ala, Arg, Asn, Asp, Glu, and Gly. The two acidic replacements mimic the effects of phosphorylation of Ser46 in HPrs from Gram-positive bacteria. In particular, when mutated to Asp46, the His 15 phosphoacceptor activity (enzyme I Km/Kcat) decreases by about 2000-fold (enzyme I Km, 4 mM HPr; Kcat, approximately 30%). The alanine and glycine mutations had near-wild-type properties, and the asparagine and arginine mutations yielded small changes to the Km values. The crystallographic tertiary structure of Ser46Asp HPr has been determined at 1.5 A resolution, and several changes have been observed which appear to be the effect of the mutation. There is a tightening of helix B, which is demonstrated by a consistent shortening of hydrogen bond lengths throughout the helix as compared to the wild-type structure. There is a repositioning of the Gly54 residue to adopt a 3(10) helical pattern which is not present in the wild-type HPr. In addition, the higher resolution of the mutant structure allows for a more definitive placement of the carbonyl of Pro11. The consequence of this change is that there is no torsion angle strain at residue 16. This result suggests that there is no active site torsion angle strain in wild-type E. coli HPr. The lack of substantial change at the active center of E. coli HPr Ser46Asp HPr suggests that the effect of the Ser46 phosphorylation in HPrs from Gram-positive bacteria is due to an electrostatic interference with enzyme I binding.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
18058890 R.Stefureac, L.Waldner, P.Howard, and J.S.Lee (2008).
Nanopore analysis of a small 86-residue protein.
  Small, 4, 59-63.  
18702519 S.Napper, L.Prasad, and L.T.Delbaere (2008).
Structural investigation of a phosphorylation-catalyzed, isoaspartate-free, protein succinimide: crystallographic structure of post-succinimide His15Asp histidine-containing protein.
  Biochemistry, 47, 9486-9496.
PDB code: 3ccd
17158705 J.Deutscher, C.Francke, and P.W.Postma (2006).
How phosphotransferase system-related protein phosphorylation regulates carbohydrate metabolism in bacteria.
  Microbiol Mol Biol Rev, 70, 939.  
16394798 J.Siebler, U.Protzer, S.Wirtz, M.Schuchmann, T.Höhler, P.R.Galle, and M.F.Neurath (2006).
Overexpression of STAT-1 by adenoviral gene transfer does not inhibit hepatitis B virus replication.
  Eur J Gastroenterol Hepatol, 18, 167-174.  
15389861 F.Meng, A.J.Forbes, L.M.Miller, and N.L.Kelleher (2005).
Detection and localization of protein modifications by high resolution tandem mass spectrometry.
  Mass Spectrom Rev, 24, 126-134.  
14690499 M.M.McKay, and R.A.Kahn (2004).
Multiple phosphorylation events regulate the subcellular localization of GGA1.
  Traffic, 5, 102-116.  
12202490 G.Cornilescu, B.R.Lee, C.C.Cornilescu, G.Wang, A.Peterkofsky, and G.M.Clore (2002).
Solution structure of the phosphoryl transfer complex between the cytoplasmic A domain of the mannitol transporter IIMannitol and HPr of the Escherichia coli phosphotransferase system.
  J Biol Chem, 277, 42289-42298.
PDB code: 1j6t
12068021 L.Athmer, J.Kindrachuk, F.Georges, and S.Napper (2002).
The influence of protein structure on the products emerging from succinimide hydrolysis.
  J Biol Chem, 277, 30502-30507.  
11551914 S.Napper, S.J.Brokx, E.Pally, J.Kindrachuk, L.T.Delbaere, and E.B.Waygood (2001).
Substitution of aspartate and glutamate for active center histidines in the Escherichia coli phosphoenolpyruvate:sugar phosphotransferase system maintain phosphotransfer potential.
  J Biol Chem, 276, 41588-41593.  
11060015 G.Wang, J.M.Louis, M.Sondej, Y.J.Seok, A.Peterkofsky, and G.M.Clore (2000).
Solution structure of the phosphoryl transfer complex between the signal transducing proteins HPr and IIA(glucose) of the Escherichia coli phosphoenolpyruvate:sugar phosphotransferase system.
  EMBO J, 19, 5635-5649.
PDB code: 1ggr
10419492 S.Napper, L.T.Delbaere, and E.B.Waygood (1999).
The aspartyl replacement of the active site histidine in histidine-containing protein, HPr, of the Escherichia coli Phosphoenolpyruvate:Sugar phosphotransferase system can accept and donate a phosphoryl group. Spontaneous dephosphorylation of acyl-phosphate autocatalyzes an internal cyclization.
  J Biol Chem, 274, 21776-21782.
PDB codes: 1cm2 1cm3
9545243 J.Dong, L.H.Hung, R.Strome, and H.M.Krause (1998).
A phosphorylation site in the ftz homeodomain is required for activity.
  EMBO J, 17, 2308-2318.  
9718298 P.P.Zhu, O.Herzberg, and A.Peterkofsky (1998).
Topography of the interaction of HPr(Ser) kinase with HPr.
  Biochemistry, 37, 11762-11770.  
9261069 P.Sliz, R.Engelmann, W.Hengstenberg, and E.F.Pai (1997).
The structure of enzyme IIAlactose from Lactococcus lactis reveals a new fold and points to possible interactions of a multicomponent system.
  Structure, 5, 775-788.
PDB code: 1e2a
8784180 R.Thapar, E.M.Nicholson, P.Rajagopal, E.B.Waygood, J.M.Scholtz, and R.E.Klevit (1996).
Influence of N-cap mutations on the structure and stability of Escherichia coli HPr.
  Biochemistry, 35, 11268-11277.  
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. Where a reference describes a PDB structure, the PDB code is shown on the right.