title: Molecular Determinants for Protein Stabilization by Insertional Fusion to a Thermophilic Host Protein.
abstract: A universal method that improves protein stability and evolution has thus far eluded discovery. Recently, however, studies have shown that insertional fusion to a protein chaperone stabilized various target proteins with minimal negative effects. The improved stability was derived from insertion into a hyperthermophilic protein, Pyrococcus furiosus maltodextrin-binding protein (PfMBP), rather than from changes to the target protein sequence. In this report, by evaluating the thermodynamic and kinetic stability of various inserted β-lactamase (BLA) homologues, we were able to examine the molecular determinants of stability realized by insertional fusion to PfMBP. Results indicated that enhanced stability and suppressed aggregation of BLA stemmed from enthalpic and entropic mechanisms. This report also suggests that insertional fusion to a stable protein scaffold has the potential to be a useful method for improving protein stability, as well as functional protein evolution.
authors.affiliation: Othmer-Jacobs Department of Chemical and Biomolecular Engineering, New York University, 6 MetroTech Center, Brooklyn, NY, 11201, USA.
authors.forename: Brennal
authors.initials: B
authors.lastname: Pierre
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, USA.
authors.forename: Jason W
authors.initials: JW
authors.lastname: Labonte
authors.affiliation: Othmer-Jacobs Department of Chemical and Biomolecular Engineering, New York University, 6 MetroTech Center, Brooklyn, NY, 11201, USA.|Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, USA.
authors.forename: Tina
authors.initials: T
authors.lastname: Xiong
authors.affiliation: Othmer-Jacobs Department of Chemical and Biomolecular Engineering, New York University, 6 MetroTech Center, Brooklyn, NY, 11201, USA.
authors.forename: Edwin
authors.initials: E
authors.lastname: Aoraha
authors.affiliation: Othmer-Jacobs Department of Chemical and Biomolecular Engineering, New York University, 6 MetroTech Center, Brooklyn, NY, 11201, USA.
authors.forename: Asher
authors.initials: A
authors.lastname: Williams
authors.affiliation: Othmer-Jacobs Department of Chemical and Biomolecular Engineering, New York University, 6 MetroTech Center, Brooklyn, NY, 11201, USA.
authors.forename: Vandan
authors.initials: V
authors.lastname: Shah
authors.affiliation: Othmer-Jacobs Department of Chemical and Biomolecular Engineering, New York University, 6 MetroTech Center, Brooklyn, NY, 11201, USA.
authors.forename: Edward
authors.initials: E
authors.lastname: Chau
authors.affiliation: Othmer-Jacobs Department of Chemical and Biomolecular Engineering, New York University, 6 MetroTech Center, Brooklyn, NY, 11201, USA.
authors.forename: Kazi Yasin
authors.initials: KY
authors.lastname: Helal
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD, 21218, USA.
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Gray
authors.affiliation: Othmer-Jacobs Department of Chemical and Biomolecular Engineering, New York University, 6 MetroTech Center, Brooklyn, NY, 11201, USA. jin.kim@nyu.edu.
authors.forename: Jin Ryoun
authors.initials: JR
authors.lastname: Kim
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002152:Calorimetry, Differential Scanning; D002850:Chromatography, Gel; D002942:Circular Dichroism; D019277:Entropy; D007700:Kinetics; D055550:Protein Stability; D058767:Protein Unfolding; D020027:Pyrococcus furiosus / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D001618:beta-Lactamases / Q000235:genetics / Q000378:metabolism
chemical_list: D019843:Archaeal Proteins; D011993:Recombinant Fusion Proteins; D001618:beta-Lactamases
keywords: protein aggregation; protein engineering; protein fusion; protein stability; protein-protein interactions