Interaction of insulin with a triblock copolymer of PEG-(fumaric-sebacic acids)-PEG: thermodynamic and spectroscopic studies.
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 x 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 beta-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 beta-structure remains unchanged. Altogether, these observations indicate that (PEG-co-P(FA/SC)-co-PEG) triblock copolymer has similar effect(s) on both proteins.
Pourhosseini
Pouneh S
PS
Institute of Biochemistry and Biophysics, University of Tehran, PO Box 13145-1384, Tehran, Iran.
Saboury
Ali A
AA
Najafi
Farhood
F
Sarbolouki
Mohammad N
MN
D000818:Animals; D002151:Calorimetry; D002417:Cattle; D002942:Circular Dichroism; D003652:Decanoic Acids / Q000737:chemistry / Q000378:metabolism*; D005650:Fumarates / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000737:chemistry / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000737:chemistry / Q000378:metabolism*; D011108:Polymers / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D017186:Titrimetry
D003652:Decanoic Acids; D005650:Fumarates; D007328:Insulin; D011108:Polymers; C473130:poly(fumaric-co-sebacic acid); D011092:Polyethylene Glycols