TY - JOUR
T1 - Structure-affinity relationship study of bleomycins and shble protein by use of a chemical array
AU - Miyazaki, Isao
AU - Okumura, Hideo
AU - Simizu, Siro
AU - Takahashi, Yoshikazu
AU - Kanoh, Naoki
AU - Muraoka, Yasuhiko
AU - Nonomura, Yoshiaki
AU - Osada, Hiroyuki
PY - 2009/3/23
Y1 - 2009/3/23
N2 - The photocrosslinked chemical array format is useful not merely for screening protein ligands, but also for gaining insight into structure-affinity relationships (SARs). By probing an array of 2000 natural products, containing 50 bleomycin (BLM) derivatives, with cell lysates that overexpress RFP-fused Shble protein, we successfully observed interactions between Shble protein and BLMs on the array. Among the BLM derivatives, those that had long C-terminal tails were found to bind strongly. The binding signal intensities observed on the chemical array correlated well with the association constants, which were determined by isothermal titration carolimetry (ITC) experiments (r2=0.663), showing that the on-chip results were not an artifact of ligand immobilization. In addition to the C-terminal tails, the propionamide moieties in pyrimidoblamic acid (PBA) also appeared to be important for binding. The contributions of the propionamide moieties of PBA to binding were further supported by the X-ray structure of the complex of Shble protein and BLM A6. These results provide insight into the structural requirements for recognition of BLMs by Shble protein.
AB - The photocrosslinked chemical array format is useful not merely for screening protein ligands, but also for gaining insight into structure-affinity relationships (SARs). By probing an array of 2000 natural products, containing 50 bleomycin (BLM) derivatives, with cell lysates that overexpress RFP-fused Shble protein, we successfully observed interactions between Shble protein and BLMs on the array. Among the BLM derivatives, those that had long C-terminal tails were found to bind strongly. The binding signal intensities observed on the chemical array correlated well with the association constants, which were determined by isothermal titration carolimetry (ITC) experiments (r2=0.663), showing that the on-chip results were not an artifact of ligand immobilization. In addition to the C-terminal tails, the propionamide moieties in pyrimidoblamic acid (PBA) also appeared to be important for binding. The contributions of the propionamide moieties of PBA to binding were further supported by the X-ray structure of the complex of Shble protein and BLM A6. These results provide insight into the structural requirements for recognition of BLMs by Shble protein.
KW - Arrays
KW - Bleomycins
KW - Photochemistry
KW - Shble protein
KW - Structure-activity relationships
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U2 - 10.1002/cbic.200800728
DO - 10.1002/cbic.200800728
M3 - Article
C2 - 19222034
AN - SCOPUS:65549104989
VL - 10
SP - 845
EP - 852
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 5
ER -