Protein Crystallization Theory Practice蛋白质结晶理论与实践.pptVIP

Protein CrystallizationTheory Practice Original document:developed by Tristan Fiedler for2003 ACA Summer School in Macromolecular Crystallography Augmented 6 July 2004 by Andy Howard Rebuilt for Biology 555, spring 2008 Journalist’s Criteria Who What When Where Why How Whither Whence (Wherefore) Who makes macromolecular crystals? Macromolecular crystals are almost always produced artificially, i.e., by human action So “who” is “scientists” What are macromolecular crystals? Crystals are translationally ordered arrays of molecules Macromolecular crystals are held together by relatively weak ionic intermolecular forces Solvent content generally above 40% When have we made them? Story goes back into the mid-19th century Systematic search for crystallization conditions dates from the 1950’s Screening kits: concept 1980’s, commercialization in 1990’s Robotics late 1990’s Nanoscale techniques early 21st century Crystallization Very old (reproduction, genetic duplication) Empirical (trial error -- ‘Screens’) ‘Art’ vs ‘Science’ Short History 1853 Hemoglobin Lehman, CG. Lehrbuch der physiologische Chemie. Leipzig 1926 Urease Sumner, JB. J Biol Chem. 69: 435 1930 Pepsin other proteolytic enzymes Northrop, JH, Kunitz, M, Herriot RM. Crystalline Enzymes. Columbia University Press, NY (review) 1934 Pepsin Diffraction Bernal JD Crowfoot, D. Nature, 133:794 1935 Tobacco Mosaic Virus Stanley, WM. Science. 81: 644 Short history (concluded) 1946: Sumner Nobel prize 1958: Myoglobin structure 1959: Hemoglobin structure 1962: Perutz/Kendrew Nobel 1979: CarterCarter paper 1985: first microgravity experiments 1990’s: commercial screening kits Late 1990’s: viable commercial crystallization robots Where do we grow them? Under mild laboratory conditions Contrast to inorganic small molecules, which are often grown from a melt Even small organic crystals often exploit temperature dependence Proteins usually avoid these techniques… Growing [Protein] Crystals Inorganic - cooli