Studies on the conformational properties of CP-10(42-55), the hinge region of CP-10, using circular dichroism and RP-HPLC.
The conformational properties of CP-10(42-55), a peptide corresponding to the hinge region of CP-10, were investigated using circular dichroism spectroscopy and reverse-phase high-performance liquid chromatography (RP-HPLC). The circular dichroism studies indicated that CP-10(42-55) formed considerable secondary structure in the presence of hydrophobic solution environments including 50% acetonitrile, 50% trifluoroethanol and 200 mM sodium dodecyl sulfate, which comprised a mixture of alpha-helix and beta-sheet. The effect of temperature on the conformation of CP-10(42-55) was investigated between 5 and 40 degrees C, with very small changes in the spectra being observed. RP-HPLC was then used to investigate the effect of temperature on the conformation of CP-10(42-55) in the presence of a hydrophobic surface. Using a C18-adsorbent, CP-10(42-55) exhibited a conformational transition at 25 degrees C, which was associated with an increase in the chromatographic contact area and the binding affinity of the peptide for the stationary phase. In addition, near-planar bandbroadening behaviour indicated that conformational species interconverted with rapid rate constants compared with the chromatographic time scale. These results indicated that the conformational change at 25 degrees C in the RP-HPLC system most likely corresponds to the unfolding of an alpha-helical and/or beta-sheet structure to an extended coil structure. Therefore, the strong chemotactic properties of this peptide may be attributed to its ability to form considerable secondary structure in the presence of a hydrophobic environment.
Lazoura
E
E
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
McLeish
M J
MJ
Aguilar
M I
MI
D020816:Amino Acid Motifs; D002135:Calcium-Binding Proteins / Q000737:chemistry*; D040501:Calgranulin A; D018925:Chemokines / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D006860:Hydrogen Bonding; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry*; D017510:Protein Folding; D017433:Protein Structure, Secondary; D009418:S100 Proteins / Q000737:chemistry*; D012996:Solutions; D013696:Temperature
D002135:Calcium-Binding Proteins; D040501:Calgranulin A; D018925:Chemokines; D010446:Peptide Fragments; D009418:S100 Proteins; C086145:S100a8 protein, mouse; D012996:Solutions