Design and solution structure of functional peptide mimetics of nerve growth factor.
The C-D loop in nerve growth factor (NGF) is involved in binding to the NGF receptor, TrkA. It is flexible and adopts several different types conformations in different NGF crystal forms. We have previously shown that a small cyclic peptide derived from the C-D loop of NGF binds to the TrkA receptor by mimicking the structure of this loop. To understand structure-function relationships in NGF C-D loop mimetics, we have produced a series of peptides predicted to form different types of beta-turns. The peptides were tested for their ability to promote cell survival in serum-free medium and to induce TrkA tyrosine phosphorylation. NMR structural studies were used to determined the backbone conformation and the spatial orientation of side chains involved in binding to the TrkA receptor. Peptides that form type I or type gammaL-alphaR beta-turns were the most active. The variety of active loop conformations suggests that the mimetics (and NGF) accommodate the binding site on TrkA by an 'induced fit' mechanism. In agreement with this hypothesis, NMR relaxation measurements detected both fast and slow motion in the peptides. We also characterized a retro-inverso peptide derived from the NGF C-D loop. This D-amino acid cyclic peptide did not adopt a conformation homologous to the NGF C-D loop and was inactive. This may be representative of difficulties in producing structural and functional mimetics by retro-inverso schemes.
Beglova
N
N
Department of Biochemistry and Montreal Joint Centre for Structural Biology, McGill University, 3655 Promenade Sir William Osler, Montréal, Québec H3G 1Y6, Canada.
Maliartchouk
S
S
Ekiel
I
I
Zaccaro
M C
MC
Saragovi
H U
HU
Gehring
K
K
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002460:Cell Line; D016895:Culture Media, Serum-Free; D015195:Drug Design; D004591:Electrophoresis, Polyacrylamide Gel; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008958:Models, Molecular; D018716:Molecular Mimicry; D020932:Nerve Growth Factor / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D010766:Phosphorylation; D017433:Protein Structure, Secondary; D051381:Rats; D020917:Receptor, trkA / Q000819:agonists / Q000737:chemistry; D012996:Solutions; D013237:Stereoisomerism; D014443:Tyrosine / Q000378:metabolism
D016895:Culture Media, Serum-Free; D010455:Peptides; D012996:Solutions; D014443:Tyrosine; D020932:Nerve Growth Factor; D020917:Receptor, trkA