title: Attenuated total reflection-FT-IR spectroscopic imaging of protein crystallization.
abstract: Protein crystallization is of strategic and commercial relevance in the post-genomic era because of its pivotal role in structural proteomics projects. Although protein structures are crucial for understanding the function of proteins and to the success of rational drug design and other biotechnology applications, obtaining high quality crystals is a major bottleneck to progress. The major means of obtaining crystals is by massive-scale screening of a target protein solution with numerous crystallizing agents. However, when crystals appear in these screens, one cannot easily know if they are crystals of protein, salt, or any other molecule that happens to be present in the trials. We present here a method based on Attenuated Total Reflection (ATR)-FT-IR imaging that reliably identifies protein crystals through a combination of chemical specificity and the visualizing capability of this approach, thus solving a major hurdle in protein crystallization. ATR-FT-IR imaging was successfully applied to study the crystallization of thaumatin and lysozyme in a high-throughput manner, simultaneously from six different solutions. This approach is fast as it studies protein crystallization in situ and provides an opportunity to examine many different samples under a range of conditions.
authors.affiliation: Department of Chemical Engineering, Faculty of Engineering, Imperial College London, SW7 2AZ, United Kingdom.
authors.forename: K L Andrew
authors.initials: KL
authors.lastname: Chan
authors.forename: Lata
authors.initials: L
authors.lastname: Govada
authors.forename: Roslyn M
authors.initials: RM
authors.lastname: Bill
authors.forename: Naomi E
authors.initials: NE
authors.lastname: Chayen
authors.forename: Sergei G
authors.initials: SG
authors.lastname: Kazarian
mesh_terms: D015703:Antigens, CD / Q000737:chemistry; D003460:Crystallization; D003461:Crystallography / Q000379:methods*; D009113:Muramidase / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D060225:Tetraspanin 28
chemical_list: D015703:Antigens, CD; D010940:Plant Proteins; D011506:Proteins; D060225:Tetraspanin 28; C003427:thaumatin protein, plant; D009113:Muramidase