TY - JOUR
T1 - Binding of peptides in solution by the Escherichia coli chaperone PapD as revealed using an inhibition ELISA and NMR spectroscopy
AU - Flemmer Karlsson, Katarina
AU - Walse, Björn
AU - Drakenberg, Torbjörn
AU - Roy, Sarbari
AU - Bergquist, Karl Erik
AU - Pinkner, Jerome S.
AU - Hultgren, Scott J.
AU - Kihlberg, Jan
N1 - Funding Information:
The authors are grateful to C. Pongratz for statistical analysis of the ELISA data for the pilus subunit derived 19-mer peptides, S. Linse for the gift of Calbindin D 28k , G. Carlström for pulse sequence programming (TRNOE) and G. Carlström and J. Evenäs for help with the 3-D-NOESY-HSQC NMR experiments. This work was funded by the Swedish National Board for Industrial and Technical Development and by the Swedish Natural Science Research Council.
PY - 1998/11
Y1 - 1998/11
N2 - PapD is the prototype member of a family of periplasmic chaperones which are required for assembly of virulence associated pili in pathogenic, gram-negative bacteria. In the present investigation, an ELISA has been developed for evaluation of compounds as inhibitors of PapD. Synthetic peptides, including an octamer, derived from the C-terminus of the pilus adhesin PapG were able to inhibit PapD in the ELISA. Evaluation of a panel of octapeptides in the ELISA, in combination with NMR studies, showed that the peptides were bound as extended β-strands by PapD in aqueous solution. The PapD-peptide complex was stabilized by backbone to backbone hydrogen bonds and interactions involving three hydrophobic peptide side chains. This structural information, together with previous crystal structure data, provides a starting point in efforts to design and synthesize compounds which bind to chaperones and interfere with pilus assembly in pathogenic bacteria. Copyright (C) 1997 Elsevier Science Ltd.
AB - PapD is the prototype member of a family of periplasmic chaperones which are required for assembly of virulence associated pili in pathogenic, gram-negative bacteria. In the present investigation, an ELISA has been developed for evaluation of compounds as inhibitors of PapD. Synthetic peptides, including an octamer, derived from the C-terminus of the pilus adhesin PapG were able to inhibit PapD in the ELISA. Evaluation of a panel of octapeptides in the ELISA, in combination with NMR studies, showed that the peptides were bound as extended β-strands by PapD in aqueous solution. The PapD-peptide complex was stabilized by backbone to backbone hydrogen bonds and interactions involving three hydrophobic peptide side chains. This structural information, together with previous crystal structure data, provides a starting point in efforts to design and synthesize compounds which bind to chaperones and interfere with pilus assembly in pathogenic bacteria. Copyright (C) 1997 Elsevier Science Ltd.
KW - Chaperone
KW - ELISA
KW - NMR spectroscopy
KW - Peptide
KW - Pili
KW - Structure-activity
UR - http://www.scopus.com/inward/record.url?scp=0031768474&partnerID=8YFLogxK
U2 - 10.1016/S0968-0896(98)00162-X
DO - 10.1016/S0968-0896(98)00162-X
M3 - Article
C2 - 9881099
AN - SCOPUS:0031768474
SN - 0968-0896
VL - 6
SP - 2085
EP - 2101
JO - Bioorganic and Medicinal Chemistry
JF - Bioorganic and Medicinal Chemistry
IS - 11
ER -