N-glycosylation of enhanced aromatic sequons to increase glycoprotein stability.
N-glycosylation can increase the rate of protein folding, enhance thermodynamic stability, and slow protein unfolding; however, the molecular basis for these effects is incompletely understood. Without clear engineering guidelines, attempts to use N-glycosylation as an approach for stabilizing proteins have resulted in unpredictable energetic consequences. Here, we review the recent development of three "enhanced aromatic sequons," which appear to facilitate stabilizing native-state interactions between Phe, Asn-GlcNAc and Thr when placed in an appropriate reverse turn context. It has proven to be straightforward to engineer a stabilizing enhanced aromatic sequon into glycosylation-naïve proteins that have not evolved to optimize specific protein-carbohydrate interactions. Incorporating these enhanced aromatic sequons into appropriate reverse turn types within proteins should enhance the well-known pharmacokinetic benefits of N-glycosylation-based stabilization by lowering the population of protease-susceptible unfolded and aggregation-prone misfolded states, thereby making such proteins more useful in research and pharmaceutical applications.
Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT 84602, USA.
Joshua L
JL
Price
Elizabeth K
EK
Culyba
Wentao
W
Chen
Amber N
AN
Murray
Sarah R
SR
Hanson
Chi-Huey
CH
Wong
Evan T
ET
Powers
Jeffery W
JW
Kelly
D000595:Amino Acid Sequence; D001665:Binding Sites; D018801:CD2 Antigens / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D008958:Models, Molecular; D011487:Protein Conformation
D018801:CD2 Antigens; D006023:Glycoproteins