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Issue 35, 2018
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Two dimensional crowding effects on protein folding at interfaces observed by chiral vibrational sum frequency generation spectroscopy

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Abstract

The crowding effect is prevalent in cellular environments due to high concentrations of biomacromolecules. It can alter the structures and dynamics of proteins and thus impact protein functions. The crowding effect is important not only in 3-dimensional cytoplasm but also for a 2-dimensional (2D) cell surface due to the presence of membrane proteins and glycosylation of membrane proteins and phospholipids. These proteins and phospholipids – with limited translational degrees of freedom along the surface normal – are confined in 2D space. Although the crowding effect at interfaces has been studied by adding crowding agents to bulk solution, the 2D crowding effect remains largely unexplored. This is mostly due to challenges in controlling 2D crowding and synergistic use of physical methods for in situ protein characterization. To address these challenges, we applied chiral vibrational sum frequency generation (SFG) spectroscopy to probe the sp1 zinc finger (ZnF), a 31-amino acid protein, folding into a β-hairpin/α-helix (ββα) motif upon binding to Zn2+. We anchored ZnF at the air/water interface via covalent linkage of ZnF to palmitic acid and controlled 2D crowding by introducing neutral lipid as a spacer. We obtained chiral amide I SFG spectra upon addition of Zn2+ and/or spacer lipid. The chiral SFG spectra show that interfacial crowding in the absence of spacer lipid hinders ZnF from folding into the ββα structure even in the presence of Zn2+. The results establish a paradigm for future quantitative, systematic studies of interfacial crowding effects.

Graphical abstract: Two dimensional crowding effects on protein folding at interfaces observed by chiral vibrational sum frequency generation spectroscopy

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Publication details

The article was received on 17 Oct 2017, accepted on 17 Aug 2018 and first published on 17 Aug 2018


Article type: Communication
DOI: 10.1039/C7CP07061F
Citation: Phys. Chem. Chem. Phys., 2018,20, 22421-22426
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    Two dimensional crowding effects on protein folding at interfaces observed by chiral vibrational sum frequency generation spectroscopy

    W. Liu, L. Fu, Z. Wang, Z. Sohrabpour, X. Li, Y. Liu, H. Wang and E. C. Y. Yan, Phys. Chem. Chem. Phys., 2018, 20, 22421
    DOI: 10.1039/C7CP07061F

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