Peptides targeting caspase inhibitors
Standard
Peptides targeting caspase inhibitors. / Tamm, Ingo; Trepel, Martin; Cardó-Vila, Marina; Sun, Yan V; Welsh, Kate; Cabezas, Edelmira; Swatterthwait, Arnold; Arap, Wadih; Reed, John C; Pasqualini, Renata.
in: J BIOL CHEM, Jahrgang 278, Nr. 16, 18.04.2003, S. 14401-5.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
Harvard
APA
Vancouver
Bibtex
}
RIS
TY - JOUR
T1 - Peptides targeting caspase inhibitors
AU - Tamm, Ingo
AU - Trepel, Martin
AU - Cardó-Vila, Marina
AU - Sun, Yan V
AU - Welsh, Kate
AU - Cabezas, Edelmira
AU - Swatterthwait, Arnold
AU - Arap, Wadih
AU - Reed, John C
AU - Pasqualini, Renata
PY - 2003/4/18
Y1 - 2003/4/18
N2 - Here we report on the identification of peptides targeting the X-inhibitor of apoptosis protein (XIAP). XIAP functions as a caspase inhibitor and is a member of the inhibitors of apoptosis (IAP) family of proteins. IAPs are often overexpressed in cancers and leukemias and are associated with an unfavorable clinical prognosis. We have selected peptides from a phage library by using recombinant full-length human XIAP or a fragment containing only the baculovirus IAP repeat 2 (BIR2) domain. A consensus motif, C(D/E/P)(W/F/Y)-acid/basic-XC, was recovered from two independent screenings by using different libraries. Phage-displaying variations of the consensus sequence bound specifically to the BIR2 domain of XIAP but not to other IAPs. The interaction was specific as it could be blocked by the cognate synthetic peptides in a dose-dependent manner. Phage displaying the XIAP-binding motif CEFESC bound to the BIR2 domain of XIAP with an estimated dissociation constant of 1.8 nm as determined by surface plasmon resonance. Protein-protein interaction assays revealed that caspase-3 and caspase-7 (but not caspase-8) blocked the binding of the CEFESC phage to XIAP, indicating that this peptide targets a domain within XIAP that is related to the caspase-binding site. In fact, the sequence EFES is homologous to a loop unique to the executioner caspase-3 and caspase-7 that are targeted by XIAP. Finally, we demonstrated that an internalizing version of the XIAP-binding peptide identified in our screenings (PFKQ) can induce programmed cell death in leukemia cells. Peptides interacting with XIAP could serve as prototypes for the design of low molecular weight modulators of apoptosis.
AB - Here we report on the identification of peptides targeting the X-inhibitor of apoptosis protein (XIAP). XIAP functions as a caspase inhibitor and is a member of the inhibitors of apoptosis (IAP) family of proteins. IAPs are often overexpressed in cancers and leukemias and are associated with an unfavorable clinical prognosis. We have selected peptides from a phage library by using recombinant full-length human XIAP or a fragment containing only the baculovirus IAP repeat 2 (BIR2) domain. A consensus motif, C(D/E/P)(W/F/Y)-acid/basic-XC, was recovered from two independent screenings by using different libraries. Phage-displaying variations of the consensus sequence bound specifically to the BIR2 domain of XIAP but not to other IAPs. The interaction was specific as it could be blocked by the cognate synthetic peptides in a dose-dependent manner. Phage displaying the XIAP-binding motif CEFESC bound to the BIR2 domain of XIAP with an estimated dissociation constant of 1.8 nm as determined by surface plasmon resonance. Protein-protein interaction assays revealed that caspase-3 and caspase-7 (but not caspase-8) blocked the binding of the CEFESC phage to XIAP, indicating that this peptide targets a domain within XIAP that is related to the caspase-binding site. In fact, the sequence EFES is homologous to a loop unique to the executioner caspase-3 and caspase-7 that are targeted by XIAP. Finally, we demonstrated that an internalizing version of the XIAP-binding peptide identified in our screenings (PFKQ) can induce programmed cell death in leukemia cells. Peptides interacting with XIAP could serve as prototypes for the design of low molecular weight modulators of apoptosis.
KW - Amino Acid Motifs
KW - Apoptosis
KW - Binding Sites
KW - Caspase 3
KW - Caspase 7
KW - Caspase 8
KW - Caspase 9
KW - Caspase Inhibitors
KW - Caspases
KW - Cell Survival
KW - Dose-Response Relationship, Drug
KW - Enzyme Inhibitors
KW - Humans
KW - Ligands
KW - Peptide Biosynthesis
KW - Peptide Library
KW - Peptides
KW - Protein Binding
KW - Protein Structure, Tertiary
KW - Proteins
KW - Surface Plasmon Resonance
KW - Time Factors
KW - X-Linked Inhibitor of Apoptosis Protein
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
KW - Research Support, U.S. Gov't, P.H.S.
U2 - 10.1074/jbc.M210133200
DO - 10.1074/jbc.M210133200
M3 - SCORING: Journal article
C2 - 12538646
VL - 278
SP - 14401
EP - 14405
JO - J BIOL CHEM
JF - J BIOL CHEM
SN - 0021-9258
IS - 16
ER -