The Ensemble of Conformations of Antifreeze Glycoproteins (AFGP8): A Study Using Nuclear Magnetic Resonance Spectroscopy

• Published in: Biomolecules
• Main Authors: Cheenou Her, Yin Yeh, Viswanathan V. Krishnan
• Format: Article in Journal/Newspaper
• Language: English
• Published: MDPI AG 2019
• Subjects: AFGP; NMR; ensemble of structures; antifreeze proteins; Microbiology; QR1-502; Antarctic; Arctic; The Antarctic; Antarc*; Arctic cod; Boreogadus saida
• Online Access: https://doi.org/10.3390/biom9060235; https://doaj.org/article/04b09a0252354b67819ec74f54fc1274

The primary sequence of antifreeze glycoproteins (AFGPs) is highly degenerate, consisting of multiple repeats of the same tripeptide, Ala−Ala−Thr*, in which Thr* is a glycosylated threonine with the disaccharide beta- d -galactosyl-(1,3)-alpha-N-acetyl- d -galactosamine. AFGPs seem to function as intrinsically disordered proteins, presenting challenges in determining their native structure. In this work, a different approach was used to elucidate the three-dimensional structure of AFGP8 from the Arctic cod Boreogadus saida and the Antarctic notothenioid Trematomus borchgrevinki . Dimethyl sulfoxide (DMSO), a non-native solvent, was used to make AFGP8 less dynamic in solution. Interestingly, DMSO induced a non-native structure, which could be determined via nuclear magnetic resonance (NMR) spectroscopy. The overall three-dimensional structures of the two AFGP8s from two different natural sources were different from a random coil ensemble, but their “compactness” was very similar, as deduced from NMR measurements. In addition to their similar compactness, the conserved motifs, Ala−Thr*−Pro−Ala and Ala−Thr*−Ala−Ala, present in both AFGP8s, seemed to have very similar three-dimensional structures, leading to a refined definition of local structural motifs. These local structural motifs allowed AFGPs to be considered functioning as effectors, making a transition from disordered to ordered upon binding to the ice surface. In addition, AFGPs could act as dynamic linkers, whereby a short segment folds into a structural motif, while the rest of the AFGPs could still be disordered, thus simultaneously interacting with bulk water molecules and the ice surface, preventing ice crystal growth.