TY  - JOUR
AU  - Tran, T.
AU  - Hoffmann, S.
AU  - Wiesehan, K.
AU  - Jonas, E.
AU  - Luge, C.
AU  - Aladag, A.
AU  - Willbold, D.
TI  - Insights into Human Lck SH3 Domain Binding Specificity: Different Binding Modes of Artificial and Native Ligands
JO  - Biochemistry
VL  - 44
SN  - 0006-2960
CY  - Columbus, Ohio
PB  - American Chemical Society
M1  - PreJuSER-49405
SP  - 15042 - 15052
PY  - 2005
N1  - Record converted from VDB: 12.11.2012
AB  - We analyzed the ligand binding specificity of the lymphocyte specific kinase (Lck) SH3 domain. We identified artificial Lck SH3 ligands using phage display. In addition, we analyzed Lck SH3 binding sites within known natural Lck SH3 binding proteins using an Lck specific binding assay on membrane-immobilized synthetic peptides. On one hand, from the phage-selected peptides, representing mostly special class I' ligands, a well-defined consensus sequence was obtained. Interestingly, a histidine outside the central polyproline motif contributes significantly to Lck SH3 binding affinity and specificity. On the other hand, we confirmed previously mapped Lck SH3 binding sites in ADAM15, HS1, SLP76, and NS5A, and identified putative Lck SH3 binding sites of Sam68, FasL, c-Cbl, and Cbl-b. Without exception, the comparatively diverse Lck SH3 binding sites of all analyzed natural Lck SH3 binding proteins emerged as class II proteins. Possible explanations for the observed variations between artificial and native ligands-which are not due to significant K(D) value differences as shown by calculating Lck SH3 affinities of artificial peptide PD1-Y(-3)R as well as for peptides comprising putative Lck SH3 binding sites of NS5A, Sos, and Sam68-are discussed. Our data suggest that phage display, a popular tool for determining SH3 binding specificity, must-at least in the case of Lck-not irrevocably mirror physiologically relevant protein-ligand interactions.
KW  - Amino Acid Sequence
KW  - Binding Sites
KW  - Consensus Sequence
KW  - Histidine: chemistry
KW  - Humans
KW  - Ligands
KW  - Lymphocyte Specific Protein Tyrosine Kinase p56(lck): chemistry
KW  - Lymphocyte Specific Protein Tyrosine Kinase p56(lck): metabolism
KW  - Peptide Library
KW  - Peptides: chemistry
KW  - Protein Binding
KW  - Sequence Alignment
KW  - Tyrosine: chemistry
KW  - src Homology Domains
KW  - Ligands (NLM Chemicals)
KW  - Peptide Library (NLM Chemicals)
KW  - Peptides (NLM Chemicals)
KW  - Tyrosine (NLM Chemicals)
KW  - Histidine (NLM Chemicals)
KW  - Lymphocyte Specific Protein Tyrosine Kinase p56(lck) (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:16274251
UR  - <Go to ISI:>//WOS:000233295900036
DO  - DOI:10.1021/bi051403k
UR  - https://juser.fz-juelich.de/record/49405
ER  -