title: Protein native-state stabilization by placing aromatic side chains in N-glycosylated reverse turns.
abstract: N-glycosylation of eukaryotic proteins helps them fold and traverse the cellular secretory pathway and can increase their stability, although the molecular basis for stabilization is poorly understood. Glycosylation of proteins at naïve sites (ones that normally are not glycosylated) could be useful for therapeutic and research applications but currently results in unpredictable changes to protein stability. We show that placing a phenylalanine residue two or three positions before a glycosylated asparagine in distinct reverse turns facilitates stabilizing interactions between the aromatic side chain and the first N-acetylglucosamine of the glycan. Glycosylating this portable structural module, an enhanced aromatic sequon, in three different proteins stabilizes their native states by -0.7 to -2.0 kilocalories per mole and increases cellular glycosylation efficiency.
authors.affiliation: Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Elizabeth K
authors.initials: EK
authors.lastname: Culyba
authors.forename: Joshua L
authors.initials: JL
authors.lastname: Price
authors.forename: Sarah R
authors.initials: SR
authors.lastname: Hanson
authors.forename: Apratim
authors.initials: A
authors.lastname: Dhar
authors.forename: Chi-Huey
authors.initials: CH
authors.lastname: Wong
authors.forename: Martin
authors.initials: M
authors.lastname: Gruebele
authors.forename: Evan T
authors.initials: ET
authors.lastname: Powers
authors.forename: Jeffery W
authors.initials: JW
authors.lastname: Kelly
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry; D017766:Acid Anhydride Hydrolases / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D001216:Asparagine / Q000737:chemistry; D018801:CD2 Antigens / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000737:chemistry; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D019696:Peptidylprolyl Isomerase / Q000737:chemistry*; D010649:Phenylalanine / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D011487:Protein Conformation; D015202:Protein Engineering; D017510:Protein Folding; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D051381:Rats; D013816:Thermodynamics; D000097024:Acylphosphatase
chemical_list: D000117:Acetylglucosamine; D017766:Acid Anhydride Hydrolases; D001216:Asparagine; D018801:CD2 Antigens; D050505:Mutant Proteins; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D019696:Peptidylprolyl Isomerase; D010649:Phenylalanine; D011134:Polysaccharides; D000097024:Acylphosphatase; C000606846:PIN1 protein, human