pH-dependent secondary conformation of the peptide hormone leptin in different buffer solutions.
The secondary structure of leptin in each different pH buffer solution (pH 5.35, 6.75, 7.58 and 8.45) was first determined by attenuated total reflection (ATR)/Fourier transform infrared (FT-IR) spectrometer with second-derivative, Fourier self-deconvolution and band curve-fitting methods to quantitatively estimate the secondary structure of leptin. The results indicate that pH induced more stretching vibration of CH2 and bending vibration of C-H and/or symmetric stretching of carboxylate of leptin structure in higher pH buffer solution than in lower pH buffer solution. Moreover, the band area of amide I for leptin in the higher pH buffer solution markedly enlarged, suggesting the amide I contour of leptin was very sensitive to pH to alter the secondary conformation of leptin structure. The structural component and composition of amide I band for leptin in both pH 6.75 and pH 7.58 buffer solutions were similar and had 50-52% helical structure including alpha-helix at 1654 cm-1 and 3(10)-helical structure at 1659-1667 cm-1 and 1640 cm-1. Although the secondary structure of leptin in pH 5.35 and 8.45 buffer solutions were also similar, a different structural information was obtained.
Au
L C
LC
Department of Medical Research and Education, Veterans General Hospital-Taipei, Taiwan.
Lin
S Y
SY
Li
M J
MJ
Ho
C J
CJ
D000818:Animals; D006728:Hormones / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D020738:Leptin; D051379:Mice; D010455:Peptides / Q000737:chemistry*; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared
D006728:Hormones; D020738:Leptin; D010455:Peptides; D011506:Proteins; D011994:Recombinant Proteins