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PDBsum entry 2ksp

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protein metals Protein-protein interface(s) links
Protein binding PDB id
2ksp
Jmol
Contents
Protein chains
105 a.a. *
15 a.a. *
Metals
_CA
* Residue conservation analysis
PDB id:
2ksp
Name: Protein binding
Title: Mechanism for the selective interaction of c-terminal eh-dom proteins with specific npf-containing partners
Structure: Eh domain-containing protein 1. Chain: a. Fragment: c-terminal domain (unp residues 435-534). Synonym: testilin, hpast1. Engineered: yes. Mical l1 like peptide. Chain: b. Synonym: molecule interacting with rab13, mirab13. Engineered: yes
Source: Homo sapiens. Human. Organism_taxid: 9606. Gene: ehd1, past, past1, cdabp0131. Expressed in: escherichia coli. Expression_system_taxid: 562. Synthetic: yes
NMR struc: 10 models
Authors: F.Kieken,M.Sharma,M.Jovic,S.S.Giridharan,N.Naslavsky,S.Capla P.L.Sorgen
Key ref: F.Kieken et al. (2010). Mechanism for the selective interaction of C-terminal Eps15 homology domain proteins with specific Asn-Pro-Phe-containing partners. J Biol Chem, 285, 8687-8694. PubMed id: 20106972
Date:
11-Jan-10     Release date:   26-Jan-10    
PROCHECK
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 Headers
 References

Protein chain
Pfam   ArchSchema ?
Q9H4M9  (EHD1_HUMAN) -  EH domain-containing protein 1
Seq:
Struc:
 
Seq:
Struc:
534 a.a.
105 a.a.*
Protein chain
Pfam   ArchSchema ?
Q8N3F8  (MILK1_HUMAN) -  MICAL-like protein 1
Seq:
Struc:
 
Seq:
Struc:
863 a.a.
15 a.a.
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 4 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Biochemical function     calcium ion binding     1 term  

 

 
J Biol Chem 285:8687-8694 (2010)
PubMed id: 20106972  
 
 
Mechanism for the selective interaction of C-terminal Eps15 homology domain proteins with specific Asn-Pro-Phe-containing partners.
F.Kieken, M.Sharma, M.Jovic, S.S.Giridharan, N.Naslavsky, S.Caplan, P.L.Sorgen.
 
  ABSTRACT  
 
Epidermal growth factor receptor tyrosine kinase substrate 15 (Eps15) homology (EH)-domain proteins can be divided into two classes: those with an N-terminal EH-domain(s), and the C-terminal Eps15 homology domain-containing proteins (EHDs). Whereas many N-terminal EH-domain proteins regulate internalization events, the best characterized C-terminal EHD, EHD1, regulates endocytic recycling. Because EH-domains interact with the tripeptide Asn-Pro-Phe (NPF), it is of critical importance to elucidate the molecular mechanisms that allow EHD1 and its paralogs to interact selectively with a subset of the hundreds of NPF-containing proteins expressed in mammalian cells. Here, we capitalize on our findings that C-terminal EH-domains possess highly positively charged interaction surfaces and that many NPF-containing proteins that interact with C-terminal (but not N-terminal) EH-domains are followed by acidic residues. Using the recently identified EHD1 interaction partner molecule interacting with CasL (MICAL)-Like 1 (MICAL-L1) as a model, we have demonstrated that only the first of its two NPF motifs is required for EHD1 binding. Because only this first NPF is followed by acidic residues, we have utilized glutathione S-transferase pulldowns, two-hybrid analysis, and NMR to demonstrate that the flanking acidic residues "fine tune" the binding affinity to EHD1. Indeed, our NMR solution structure of the EHD1 EH-domain in complex with the MICAL-L1 NPFEEEEED peptide indicates that the first two flanking Glu residues lie in a position favorable to form salt bridges with Lys residues within the EH-domain. Our data provide a novel explanation for the selective interaction of C-terminal EH-domains with specific NPF-containing proteins and allow for the prediction of new interaction partners with C-terminal EHDs.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
20961375 B.Cai, D.Katafiasz, V.Horejsi, and N.Naslavsky (2011).
Pre-sorting endosomal transport of the GPI-anchored protein, CD59, is regulated by EHD1.
  Traffic, 12, 102-120.  
21067929 N.Naslavsky, and S.Caplan (2011).
EHD proteins: key conductors of endocytic transport.
  Trends Cell Biol, 21, 122-131.  
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