An Overview of Biological Macromolecule Crystallization

Abstract

The elucidation of the three dimensional structure of biological macromolecules has provided an important contribution to our current understanding of many basic mechanisms involved in life processes. This enormous impact largely results from the ability of X-ray crystallography to provide accurate structural details at atomic resolution that are a prerequisite for a deeper insight on the way in which bio-macromolecules interact with each other to build up supramolecular nano-machines capable of performing specialized biological functions. With the advent of high-energy synchrotron sources and the development of sophisticated software to solve X-ray and neutron crystal structures of large molecules, the crystallization step has become even more the bottleneck of a successful structure determination. This review introduces the general aspects of protein crystallization, summarizes conventional and innovative crystallization methods and focuses on the new strategies utilized to improve the success rate of experiments and increase crystal diffraction quality.

Native-like Photosystem II Superstructure at 2.44 Å Resolution through Detergent Extraction from the Protein Crystal

“…A detergent is needed to extract the protein from the membrane, whereupon a detergent belt around the hydrophobic surface is formed (Garavito and Ferguson-Miller, 2001). This belt is not incorporated into type I crystals, where layers of membrane proteins are stacked with protein contacts between transmembrane regions (Russo Krauss et al, 2013;Michel, 1983). In contrast, the space between protein units in type II crystals is usually large enough to accommodate the original detergent belt at the expense of stable protein contacts, which likely contributes to the difficulty in obtaining high-quality crystals for diffraction experiments.…”

“…A prominent method among postcrystallization treatments to improve crystal quality is controlled dehydration (Russo Krauss et al, 2013;Heras and Martin, 2005). This method was used by Umena et al (2011) to improve dPSIIcc crystals from T. vulcanus, using the detergent combination bDM/HTG, and ultimately yielded a resolution of 1.9 Å .…”

Identification of Anticancer Lead Molecules against PRR11 Protein Target with Combination of Protein Modelling Through Threading Approach, Structure Based Chemical Screening of ZINC Database and Pharmacokinetic Properties

“…The protein sequences deposited in UniProt is above 20 million but their crystal structures deposited in PDB is only 90,000, this give us a clear idea that the challenge in determining crystal structures of proteins is arduous. 8 The protein targeted chemo therapy being played a vital role in the treatment of diseases. Due to insufficient crystal structures of target proteins, the bioinformatics tools taken a step to build 3D models based on homology of template crystal structures of others.…”

“…Hence, target protein atomic and molecular structure through X-ray crystallography and NMR, further identification of drugs against such targets is urgent necessary. 7,8 Determination of protein structures through experimental methods are tended to very slow and not succeed for all proteins especially for membrane proteins. 9 Bioinformatics is a new discipline emerged from engage of biology and computer science, it is a site of address for many complicated questions which are not answered by experimental methods.…”

Estudos estruturais de enzimas da via de biossíntese de folatos de <i>Xanthomonas albilineans</i> para o desenvolvimento de novos candidatos a agroquímicos para a cultura de cana-de-açúcar

“…Esse evento inicial e fundamental para obtenção de um cristal de proteína é denominado nucleação. [131][132][133][134][135] Seguido da nucleação, a difusão das proteínas em solução para a superfície do cristal leva ao crescimento ordenado dos núcleos formados, em um evento denominado fase de crescimento, como mostrado no diagrama de fases de cristalização (Figura 25). A formação da rede cristalina depende fortemente de interações especificas e espacialmente orientadas entre as proteínas ainda em solução e os núcleos críticos, fazendo deste um evento dependente da homogeneidade oligomérica da amostra de proteína.…”