TY  - JOUR
AU  - Bauer, F.
AU  - Hofinger, E.
AU  - Hoffmann, S.
AU  - Rösch, P.
AU  - Schweimer, K.
AU  - Sticht, H.
TI  - Characterization of Lck-binding elements in the herpesviral regulatory Tip protein
JO  - Biochemistry
VL  - 43
SN  - 0006-2960
CY  - Columbus, Ohio
PB  - American Chemical Society
M1  - PreJuSER-46280
SP  - 14932-9
PY  - 2004
N1  - Record converted from VDB: 12.11.2012
AB  - Herpesvirus saimiri encodes a tyrosine kinase interacting protein (Tip) that binds to T-cell-specific tyrosine kinase Lck via multiple sequence motifs and controls its activity. The regulation of Lck by Tip represents a key mechanism in the transformation of human T-lymphocytes during herpesviral infection. In this study, the interaction of Tip with the regulatory SH3 and SH2 domains of Lck was investigated by biophysical and computational techniques. NMR spectroscopy of isotopically labeled Tip(140-191) revealed that the interaction with the LckSH3 domain is not restricted to the classical proline-rich motif, but also involves the C-terminally adjacent residues which pack into a hydrophobic pocket on the surface of the SH3 domain, thus playing a likely role in mediating binding specificity. Fluorescence binding studies of Tip further demonstrate that Tyr127 in its phosphorylated form represents a strong ligand of the LckSH2 domain, indicating the presence of an additional Lck interaction motif. In contrast, Tyr114, known to be essential for STAT-3 binding, does not interact with the LckSH2 domain, showing that the tyrosines in Tip exhibit distinct binding specificity. The existence of numerous interaction sites between Tip and the regulatory domains of Lck implies a complex regulatory mechanism and may have evolved to allow a gradual regulation of Lck activity in different pathogenic states.
KW  - Amino Acid Sequence
KW  - Circular Dichroism
KW  - Cloning, Molecular
KW  - Conserved Sequence
KW  - Herpesvirus 2, Saimiriine: chemistry
KW  - Herpesvirus 2, Saimiriine: enzymology
KW  - Herpesvirus 2, Saimiriine: metabolism
KW  - Hydrophobic and Hydrophilic Interactions
KW  - Kinetics
KW  - Ligands
KW  - Lymphocyte Specific Protein Tyrosine Kinase p56(lck): chemistry
KW  - Lymphocyte Specific Protein Tyrosine Kinase p56(lck): metabolism
KW  - Models, Molecular
KW  - Molecular Sequence Data
KW  - Nuclear Magnetic Resonance, Biomolecular
KW  - Phosphoproteins: chemistry
KW  - Phosphoproteins: genetics
KW  - Phosphoproteins: isolation & purification
KW  - Phosphoproteins: metabolism
KW  - Phosphotyrosine: chemistry
KW  - Protein Binding
KW  - Protein Structure, Secondary
KW  - Protons
KW  - Sequence Homology, Amino Acid
KW  - Spectrometry, Fluorescence
KW  - Viral Proteins: chemistry
KW  - Viral Proteins: genetics
KW  - Viral Proteins: isolation & purification
KW  - Viral Proteins: metabolism
KW  - Ligands (NLM Chemicals)
KW  - Phosphoproteins (NLM Chemicals)
KW  - Protons (NLM Chemicals)
KW  - Viral Proteins (NLM Chemicals)
KW  - tyrosine kinase interacting protein, Saimiriine herpesvirus 2 (NLM Chemicals)
KW  - Phosphotyrosine (NLM Chemicals)
KW  - Lymphocyte Specific Protein Tyrosine Kinase p56(lck) (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:15554700
UR  - <Go to ISI:>//WOS:000225353300007
DO  - DOI:10.1021/bi0485068
UR  - https://juser.fz-juelich.de/record/46280
ER  -