Retinol and retinoic acid bind human serum albumin: stability and structural features.
Vitamin A components, retinol and retinoic acid, are fat-soluble micronutrients and critical for many biological processes, including vision, reproduction, growth, and regulation of cell proliferation and differentiation. The cellular uptake of Vitamin A is through specific interaction of a plasma membrane receptor with serum retinol-binding protein. Human serum albumin (HSA), as a transport protein, is the major target of several micronutrients in vivo. The aim of present study was to examine the interaction of retinol and retinoic acid with human serum albumin in aqueous solution at physiological conditions using constant protein concentration and various retinoid contents. FTIR, UV-vis, CD and fluorescence spectroscopic methods were used to determine retinoid binding mode, the binding constant and the effects of complexation on protein secondary structure. Structural analysis showed that retinol and retinoic acid bind non-specifically (H-bonding) via protein polar groups with binding constants of K(ret)=1.32 (+/-0.30)x10(5)M(-1) and K(retac)=3.33 (+/-0.35)x10(5)M(-1). The protein secondary structure showed no alterations at low retinoid concentrations (0.125 mM), whereas at high retinoid content (1mM), an increase of alpha-helix from 55% (free HSA) to 60% and a decrease of beta-sheet from 22% (free HSA) to 18% occurred in the retinoid-HSA complexes. The results point to a partial stabilization of protein secondary structure at high retinoid content.
N'soukpoé-Kossi
C N
CN
Department of Chemistry-Biology, Université du Québec à Trois-Riviéres, C.P. 500, Trois-Riviéres, Québec, Canada G9A 5H7.
Sedaghat-Herati
R
R
Ragi
C
C
Hotchandani
S
S
Tajmir-Riahi
H A
HA
D006801:Humans; D007700:Kinetics; D011485:Protein Binding; D017433:Protein Structure, Secondary; D012176:Retinoids / Q000737:chemistry / Q000378:metabolism; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D017550:Spectroscopy, Fourier Transform Infrared; D013816:Thermodynamics; D014212:Tretinoin / Q000737:chemistry* / Q000378:metabolism*; D014801:Vitamin A / Q000737:chemistry* / Q000378:metabolism*
D012176:Retinoids; D012709:Serum Albumin; D014801:Vitamin A; D014212:Tretinoin