Disordered nano-wrinkle substrates for inducing crystallization over a wide range of concentration of protein and precipitant

TL;DR

This paper introduces nano-wrinkled substrates that significantly enhance protein crystallization across a wide range of concentrations, even without precipitants, offering a universal tool for crystallizing difficult proteins.

Contribution

The study presents a novel nano-wrinkled substrate that improves protein crystallization efficiency and control, reducing dependence on specific conditions and precipitants.

Findings

Crystals nucleate at very low protein concentrations.

Crystals form over a wide precipitant concentration range (~0.003-0.3M).

Crystals can form without any precipitant for some proteins.

Abstract

There are large number of proteins, the existence of which are known but not their crystal structure, because of difficulty in finding the exact condition for their crystallization. Heterogeneous nucleation on disordered porous substrates with small yet large distribution of pores is considered a panacea for this problem, but a universal nucleant, suitable for crystallizing large variety of proteins does not really exist. To this end, we report here a nano-wrinkled substrate which displays remarkable ability and control over protein crystallization. Experiments with different proteins show that on these substrates, crystals nucleate even at very low protein concentration in buffer. Small number of very large crystals appear for precipitant concentrations varied over orders of magnitude ~0.003-0.3M; for some proteins, crystals appear even without addition of any precipitant, not seen with any other heterogeneous substrates. In essence, these substrates significantly diminish the influence of the above two parameters, thought to be key factor for crystallization, signifying that this advantage can be exploited for finding out crystallization condition for other yet-to-be-crystallized proteins.