CHAPTER FIVE Advances in Nanocrystallography as a Proteomic Tool Eugenia Pechkova * ,† , Nicola Luigi Bragazzi * ,†,{ , Claudio Nicolini * ,†,},1 * Nanobiotechnology and Biophysics Laboratories (NBL), Department of Experimental Medicine (DIMES), University of Genoa, Genoa, Italy † Nanoworld Institute Fondazione ELBA Nicolini (FEN), Pradalunga, Bergamo, Italy { School of Public Health, Department of Health Sciences (DISSAL), University of Genoa, Genoa, Italy } Biodesign Institute, Arizona State University, Tempe, Arizona, USA 1 Corresponding author: e-mail address: info@fondazioneelba-nicolini.org Contents 1. Introduction 164 2. Langmuir–Blodgett (LB)-Based Crystallization 166 3. Comparison of LB-Based Crystallization with Other Techniques 168 4. Fourier Transform Infrared (FTIR) Spectroscopy for Investigating LB-Films 170 5. Raman Spectroscopy 171 6. Laser-Induced Microdissection and Microfragmentation 172 7. Micrograzing-Incidence X-Ray Scattering Angle (m-GISAXS) 172 8. In Silico Simulations 176 9. Bioinformatics 177 10. Molecular Dynamics 178 11. Clinically Relevant Proteins 180 11.1 GroEL 180 11.2 Casein kinase 2 181 11.3 Cytochrome P-450 side-chain cleavage 181 11.4 Rhodopsin 182 11.5 Globins 183 11.6 Insulin 183 12. Conclusions 184 References 185 Abstract In order to overcome the difficulties and hurdles too much often encountered in crys- tallizing a protein with the conventional techniques, our group has introduced the inno- vative Langmuir–Blodgett (LB)-based crystallization, as a major advance in the field of both structural and functional proteomics, thus pioneering the emerging field of the so-called nanocrystallography or nanobiocrystallography. This approach uniquely com- bines protein crystallography and nanotechnologies within an integrated, coherent Advances in Protein Chemistry and Structural Biology, Volume 95 # 2014 Elsevier Inc. ISSN 1876-1623 All rights reserved. http://dx.doi.org/10.1016/B978-0-12-800453-1.00005-1 163