title: Hydrophobic Interaction Between Domain I of Albumin and B Chain of Detemir May Support Myristate-Dependent Detemir-Albumin Binding.
abstract: The bindings of detemir [LysB29(Nε-tetradecanoyl)des(B30)-insulin] with two highly homologous albumins, HSA (human serum albumin) and BSA (bovine serum albumin), were investigated through CD, spectrofluorophotometry, and molecular docking analysis. The absence of any tryptophanyl residue in detemir makes albumin binding study possible by exclusive tryptophanyl spectral quenching at 340 nm (λem = 296 nm). The interactions found to be static (Kq > 1010M-1s-1) with Stern-Volmer constants ≈103M-1. The observed ΔG0that was negative in all cases concludes the reactions were spontaneous. Domains I and III of an albumin unfold with 5.0 M urea at pH 7.4, although domain II remains intact. Significant decreases in ΔH0and ΔS0were due to unfolding explicit that detemir binding may involve domains I and III of albumins. Temperature-dependent changes in binding were higher in HSA than BSA but after unfolding such changes were very less, further indicating the role of domains I and III in detemir binding. Pro28 and Tyr26 of insulin were found to be interacting with Arg114 and Val116 of HSA domain I, while myristate segment of detemir binds to Lys519 of domain III. Interactions seem to be predominantly hydrophobic and entropy driven. Although detemir binds to albumin through myristate, the peptide part shows involvement in binding.
authors.affiliation: Genes and Proteins Lab, Department of Biotechnology, Jamia Millia Islamia, New Delhi, 110025, India. sfatima1@jmi.ac.in.
authors.forename: Sadaf
authors.initials: S
authors.lastname: Fatima
authors.affiliation: Centre for Bioseparation Technology, VIT University, Vellore, 632014, India.
authors.forename: Priyankar
authors.initials: P
authors.lastname: Sen
authors.affiliation: Department of Integrative Biology, School of Bio Sciences and Technology, VIT University, Vellore, 632014, India.
authors.forename: P
authors.initials: P
authors.lastname: Sneha
authors.affiliation: Department of Integrative Biology, School of Bio Sciences and Technology, VIT University, Vellore, 632014, India.
authors.forename: C George
authors.initials: CG
authors.lastname: Priyadoss
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D000069057:Insulin Detemir / Q000737:chemistry*; D007700:Kinetics; D062105:Molecular Docking Simulation; D019814:Myristic Acid / Q000737:chemistry*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D058767:Protein Unfolding; D012709:Serum Albumin / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013816:Thermodynamics
chemical_list: D012709:Serum Albumin; D019814:Myristic Acid; D012710:Serum Albumin, Bovine; D000069057:Insulin Detemir
keywords: Binding thermodynamics; Bovine serum albumin; Detemir; Human serum albumin; Protein-ligand interaction; Stern–Volmer quenching