title: Expression of a human serum albumin fragment (consisting of subdomains IA, IB, and IIA) and a study of its properties.
abstract: Site-directed mutagenesis and a yeast expression system were used to synthesize a human serum albumin (HSA) fragment (amino acids 1-297). The HSA fragment (half HSA) was evaluated with a number of biophysical techniques and found to be similar to the corresponding region in wild-type HSA. Specifically, the circular dichroism spectra of half HSA and wild-type HSA were superimposable, indicating that the highly alpha-helical secondary structure of wild-type HSA is preserved in half HSA. Additionally, half HSA was partially reactive with a polyclonal antibody against authentic HSA. Half HSA, which contains subdomain IIA, had an affinity for thyroxine and several thyroxine analogs, similar to that observed previously for wild-type HSA. This study suggests that the production of recombinant HSA fragments will be useful for the study of HSA ligand interactions.
authors.affiliation: Department of Biochemistry and Biophysics, John A. Burns School of Medicine, University of Hawaii, Honolulu 96822, USA.
authors.forename: D S
authors.initials: DS
authors.lastname: Park
authors.forename: C E
authors.initials: CE
authors.lastname: Petersen
authors.forename: C
authors.initials: C
authors.lastname: Ha
authors.forename: K
authors.initials: K
authors.lastname: Harohalli
authors.forename: J B
authors.initials: JB
authors.lastname: Feix
authors.forename: N V
authors.initials: NV
authors.lastname: Bhagavan
mesh_terms: D055592:Biophysical Phenomena; D001703:Biophysics; D002942:Circular Dichroism; D004578:Electron Spin Resonance Spectroscopy; D015870:Gene Expression; D006801:Humans; D007700:Kinetics; D008024:Ligands; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D010843:Pichia / Q000235:genetics; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012709:Serum Albumin / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D013974:Thyroxine / Q000031:analogs & derivatives / Q000378:metabolism
chemical_list: D008024:Ligands; D010446:Peptide Fragments; D011994:Recombinant Proteins; D012709:Serum Albumin; D013974:Thyroxine