Interaction of insulin with a triblock copolymer of PEG-(fumaric-sebacic acids)-PEG: Thermodynamic and spectroscopic studies

• Published in: Biochimica et biophysica acta 2007-10, Vol.1774 (10), p.1274-1280
• Main Authors: Pourhosseini, Pouneh S., Saboury, Ali A., Najafi, Farhood, Sarbolouki, Mohammad N.
• Format: Article
• Language: English
• Subjects: Acrylamide quenching; Animals; Calorimetry; Cattle; Circular Dichroism; Copolymers; Decanoic Acids - chemistry; Decanoic Acids - metabolism; Denaturation; Fumarates - chemistry; Fumarates - metabolism; Humans; Insulin; Insulin - chemistry; Insulin - metabolism; ITC; Polyethylene Glycols - chemistry; Polyethylene Glycols - metabolism; Polymers - chemistry; Polymers - metabolism; Spectrometry, Fluorescence; Thermodynamics; Titrimetry
• ISSN: 1570-9639; 0006-3002; 1878-1454
• DOI: 10.1016/j.bbapap.2007.08.007

A comparative study on the interaction of (PEG-co-P(FA/SC)-co-PEG) triblock copolymer with bovine and human insulins was carried out using isothermal titration calorimetry (ITC), circular dichroism (CD), and fluorescence spectroscopy. ITC data show that the copolymer has a low affinity for both proteins, with an association constant of about 7–9 × 10 3 M − 1 . Results also show that binding is enthalpically driven, and disfavored by conformational entropy. CD spectroscopy studies reveal a small increase in the helical content and a decrease in β-structure as well as random coil in both proteins. Acrylamide quenching experiments display reduced accessibility of tyrosines, while intrinsic fluorescence spectra show lower tyrosine emission. Furthermore, thermal unfolding experiments, studied by far-UV CD at 222 and 217 nm, demonstrate that upon interaction with the copolymer helix structure becomes less stable while the stability of β-structure remains unchanged. Altogether, these observations indicate that (PEG-co-P(FA/SC)-co-PEG) triblock copolymer has similar effect(s) on both proteins.