Macromolecular crystallization: basics and advanced methodologies
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REVIEW
Macromolecular crystallization: basics and advanced methodologies Hamid Cheraghian Radi1 · Behnam Hajipour‑Verdom2 · Fatemeh Molaabasi3 Received: 11 July 2020 / Accepted: 3 September 2020 © Iranian Chemical Society 2020
Abstract For the first time, about 155 years ago and as a laboratory curiosity, German biologists observed the crystals of hemoglobin from worms and fishes. Since then, the crystallization of proteins, nucleic acids and big biological structures, like viruses, has been developed into a broad research field including several applications, for example in the drug discovery. This review is divided into four major sections. The first section addresses the specific physicochemical properties of biomolecular crystals accompanied by kinetics of supersaturation, nucleation and growth which are the three main steps required to achieve macromolecular crystals. Besides, various physical, chemical and biochemical parameters influencing the process of macromolecular crystallization are reviewed. The second part deals with classical approaches, such as vapor and batch diffusion methods, available to create macromolecular crystals. The third part overviews novel approaches including microgravity, cocrystallization, membrane-assisted crystallization and microfluidic array chips involved in more complicated techniques for growing macromolecular crystals and controlling their size and orientation. In the end, considering the very significant role of automation in providing biomolecules’ crystals in recent years, we provided a brief explanation about robotics and their importance in developing high-throughput crystallization. RDC-NMR and SAXS/WAXS hybrid methods with the aim of obtaining structural information of complex macromolecular assemblies are also discussed. Keywords Crystallization · Biomacromolecule · Supersaturation · Nucleation
Introduction Structural biology grabbed huge attention in the past few years because it plays a major role in both structure-based drug design (SBDD) programs in the pharmaceutical industry and structural genomics programs. Today, there are two main physicochemical approaches for determining the 3D structure of a macromolecule: NMR spectroscopy and X-ray crystallography [1]. While the former technique with short transverse relaxation times, chemical shift overlap and enhanced spectral complexity is capable to resolve the
* Fatemeh Molaabasi [email protected] 1
Department of Microbial Biotechnology, School of Biology and Center of Excellence in Phylogeny of Living Organisms, College of Science, University of Tehran, Tehran, Iran
2
Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran 14115‑154, Iran
3
Department of Biomaterials and Tissue Engineering, Breast Cancer Research Center, Motamed Cancer Institute, ACECR, Tehran 15179‑64311, Iran
atomic structure of macromolecules with an upper molecular weight limit of approximately 25 kDa, or approximately 220 amino acids, the latter one is better for determining
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