Gas phase dimerization of neuropeptide head activator analogs useful for the noncovalent constraint of peptides.
We have used electrospray mass spectrometry to examine the dimerization of EFLIVKS, a reversed sequence analog of part of the neuropeptide head activator, and other similar analogs. Observation of the EFLIVKS gas phase dimer is concentration-dependent, with a half-saturation concentration for relative dimer formation of 7.8 microM, similar to that of SKVILFE of 12 microM. The lowest energy conformers from quenched molecular dynamics simulations suggest EFLIVKS may dimerize in the gas phase by formation of multiple ion pairs across the dimer interface. Alanine-scan mutants also dimerize in the gas phase, with replacement of the interior residues FLIVK diminishing dimerization. The concentration-dependence of the EFLIVKS circular dichroism spectrum at pH 7.5 suggests the existence of different conformation states at different concentrations, but does not provide evidence supporting the saturable dimer formation in solution. Different analogs of EFLIVKS, when fused to each end of a 18mer unfolded peptide, induce solution structures with T(m)s of 42-50 degrees C. These peptides and analogs may thus be useful for the noncovalent constraint of peptides and peptide library members used in cellular screens.
Gururaja
Tarikere L
TL
Rigel Inc., 1180 Veterans Blvd., South San Francisco, CA 94080, USA.
Payan
Donald G
DG
Anderson
D C
DC
D000595:Amino Acid Sequence; D001704:Biopolymers / Q000737:chemistry; D002942:Circular Dichroism; D019281:Dimerization; D005740:Gases; D008958:Models, Molecular; D009479:Neuropeptides / Q000737:chemistry*; D009842:Oligopeptides / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry; D019151:Peptide Library; D011487:Protein Conformation; D020836:Protein Structure, Quaternary; D012996:Solutions; D021241:Spectrometry, Mass, Electrospray Ionization; D013816:Thermodynamics
D001704:Biopolymers; D005740:Gases; D009479:Neuropeptides; D009842:Oligopeptides; D010446:Peptide Fragments; D019151:Peptide Library; D012996:Solutions