Microsystems for UV-visible and x-ray analysis of protein crystals

L. Cheung, R. Quick, S.K. Singh, A. Weichsel, W. Montfort, Y. Zohar
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Abstract

polydimethylsiloxane (PDMS) based microsystems have successfully been fabricated and characterized for studying protein crystals utilizing both UV-visible spectroscopy and X-ray crystallography. Transmittance tests have been conducted with PDMS and glass substrates; the measurements indicate that in PDMS, unlike glass, the emerging intensity is higher than 50% of the incident intensity as long as the total optical path is shorter than 100 mum. Indeed, both the UV-visible spectrum and X-ray diffraction of a protein crystal enclosed in a PDMS device are almost identical to those of the crystal alone. Hence, PDMS is suitable as substrate material in device fabrication to study protein crystals. In glass, however, the UV-visible spectrum is significantly distorted and the X-ray diffraction pattern is rather weak resulting in poor signal to noise ratio. Furthermore, microsystems integrated with micro- channels allowing continuous exchange of buffer solution around the protein crystals have been tested; this would greatly enhance the potential to induce, trap and characterize functional states in proteins.
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用于蛋白质晶体紫外可见和x射线分析的微系统
基于聚二甲基硅氧烷(PDMS)的微系统已成功制备,并利用紫外可见光谱和x射线晶体学对其进行了表征。在PDMS和玻璃基板上进行了透光率测试;测量结果表明,在PDMS中,与玻璃不同,只要总光程短于100 μ m,新兴强度高于入射强度的50%。事实上,包裹在PDMS装置中的蛋白质晶体的紫外可见光谱和x射线衍射几乎与单独的晶体相同。因此,PDMS适合作为研究蛋白质晶体的器件制造中的衬底材料。然而,在玻璃中,紫外可见光谱明显扭曲,x射线衍射图样较弱,导致信噪比较差。此外,与微通道集成的微系统允许在蛋白质晶体周围连续交换缓冲溶液,已经进行了测试;这将极大地增强诱导、捕获和表征蛋白质功能状态的潜力。
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