Mix-and-extrude: high-viscosity sample injection towards time-resolved protein crystallography

IF 6.1 3区材料科学 Q1 Biochemistry, Genetics and Molecular Biology Journal of Applied Crystallography Pub Date : 2023-05-12 DOI:10.1101/2022.11.23.517685

M. Vakili, Huijong Han, C. Schmidt, A. Wrona, M. Kloos, I. de Diego, K. Dörner, T. Geng, Chan Kim, F. Koua, D. Melo, M. Rappas, A. Round, E. Round, M. Sikorski, J. Valerio, T. Zhou, K. Lorenzen, J. Schulz

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引用次数: 1

Abstract

Time-resolved crystallography enabled the visualization of protein molecular motion during reaction. While light is commonly used to initiate reactions in time-resolved crystallography, only a small number of proteins can in fact be activated by light. However, many biological reactions can be triggered by the interaction of proteins with ligands. The sample delivery method presented here uses a mix-and-extrude approach based on 3D printed microchannels in conjunction with a micronozzle to study the dynamics of samples in viscous media that can be triggered by diffusive mixing. The device design allows for mixing of ligands and protein crystals in a time window of 2 to 20 seconds. The device characterization using a model system (fluorescence quenching of iq-mEmerald proteins by copper ions) demonstrated that ligand and protein crystals, each within the lipidic cubic phase, can be mixed efficiently. The potential use of this approach for time-resolved membrane protein crystallography to support in the development of new drugs is also discussed. Synopsis 3D printed mixing-HVE devices address time-resolved membrane protein crystallography challenges via compact dual-flow LCP injection.

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混合和挤出：高粘度样品注入时间分辨蛋白质晶体学

时间分辨晶体学使蛋白质分子在反应过程中的运动可视化。虽然在时间分辨晶体学中，光通常用于引发反应，但事实上只有少数蛋白质可以被光激活。然而，蛋白质与配体的相互作用可以引发许多生物反应。本文提出的样品递送方法使用基于3D打印微通道的混合和挤出方法，结合微喷嘴，研究扩散混合可能引发的粘性介质中样品的动力学。该装置设计允许在2至20秒的时间窗口内混合配体和蛋白质晶体。使用模型系统的器件表征（铜离子对iq-mEmerald蛋白的荧光猝灭）表明，配体和蛋白晶体（均在脂质立方相中）可以有效混合。还讨论了这种方法在时间分辨膜蛋白晶体学中的潜在用途，以支持新药的开发。简介3D打印混合HVE设备通过紧凑的双流LCP注射解决了时间分辨膜蛋白晶体学的挑战。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Applied Crystallography 化学-晶体学

CiteScore

10.00

自引率

3.30%

发文量

178

审稿时长

4.7 months

期刊介绍： Many research topics in condensed matter research, materials science and the life sciences make use of crystallographic methods to study crystalline and non-crystalline matter with neutrons, X-rays and electrons. Articles published in the Journal of Applied Crystallography focus on these methods and their use in identifying structural and diffusion-controlled phase transformations, structure-property relationships, structural changes of defects, interfaces and surfaces, etc. Developments of instrumentation and crystallographic apparatus, theory and interpretation, numerical analysis and other related subjects are also covered. The journal is the primary place where crystallographic computer program information is published.