Femtosecond Laser Processing of Agarose Gel Surrounding Protein Crystals for Development of an Automated Crystal Capturing System

スポンサーリンク

概要

• 論文の詳細を見る

• Protein crystals must be captured to be mounted onto the goniometer head of X-ray diffraction equipment for structural analysis. However, this capturing operation has to be performed manually under microscopic observation. Crystallographers often face problems with this operation because protein crystals are very soft and fragile. Here, we crystallized elastase, thaumatin, glucose isomerase, and lysozyme in 2.0% (w/v) agarose gels and applied a femtosecond laser to process the agarose gel surrounding the protein crystals. A software-based operation system was established to enable automated laser processing. This new approach allows high-speed, high-precision, and reproducible processing of the gel without unsealing the crystallization trays. The processed gel containing crystals could be captured using a nylon loop without difficulty, followed by mounting the crystal onto the goniometer head of the X-ray diffraction equipment. X-ray diffraction analysis of such crystals suggested that the processed agarose gel with a thickness of approximately ${<}0.2$ mm has little effect on the background X-ray scattering. Furthermore, the effect of laser irradiation was investigated by X-ray diffraction and subsequent structural analyses, which demonstrated that the quality of the diffraction data and obtained electron density was essentially the same as that obtained before laser irradiation. On the other hand, the manually processed gel-grown crystals gave higher values on the background X-ray scattering. These comparative experimental results show clear advantages of our laser processing system. This approach leads to the possibility that protein crystals can be captured reproducibly without affecting any later crystallographic analysis, thereby providing an automated system for crystal capture.

• 2009-10-25

著者

• Sugiyama Shigeru

  Graduate School Of Engineering Osaka University

• Takano Kazufumi

  Graduate School Of Engineering Osaka University:sosho Inc.:crest Jst

• Kitatani Tomoya

  Graduate School Of Engineering Osaka University

• Adachi Hiroaki

  Graduate School Of Engineering Osaka University:sosho Inc.:crest Jst

• Hirose Mika

  Graduate School Of Engineering Osaka University

• Inoue Tsuyoshi

  Graduate School Of Biostudies Kyoto University

• Matsumura Hiroyoshi

  Graduate School Of Engineering Osaka University

• Mori Yusuke

  Graduate School Of Bionics Tokyo University Of Technology

• Shimizu Noriko

  Graduate School Of Engineering Osaka University

• Takahashi Yoshinori

  Graduate School And Faculty Of Science Himeji Institute Of Technology

• Kitatani Tomoya

  Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan

• Hasenaka Hitoshi

  Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan

• Murakami Satoshi

  SOSHO Inc., Osaka 541-0053, Japan

• Sugiyama Shigeru

  Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan

関連論文

• EP-P36 Structural and functional characterization of glycerol kinase from the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1(Section VII Enzyme and Protein Engineering)
• Proline Effect on the Thermostability and Slow Unfolding of a Hyperthermophilic Protein
• 2P-078 大腸菌由来pitrilysinの安定性解析(蛋白質・物性(2),第46回日本生物物理学会年会)
• Protein Thermostabilization Requires a Fine-tuned Placement of Surface-charged Residues
• Application of Femtosecond Laser Ablation for Detaching Grown Protein Crystals from Glass Capillary Tube(METHODS)
• 3F11-3 Crystal structures of cavity-filling mutants of E. coli RNase HI
• 2P-119 ヒトIgGに結合するRNAアプタマー複合体の立体構造(核酸・相互作用,複合体,第46回日本生物物理学会年会)
• New Technique of Manipulating a Protein Crystal Using Adhesive Material
• Solution-Stirring Method Improves Crystal Quality of Human Triosephosphate Isomerase(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
• Processing of Membrane Protein Crystal Using Ultraviolet Laser Irradiation(METHODS, Advances in Biomedical Science and Engineering)
• Femtosecond Laser Processing of Protein Crystals in Crystallization Drop
• Protein Cryocrystallography Using Laser-Processed Crystal
• Membrane Protein Crystallization Using Laser Irradiation
• 2P007 大腸菌由来スペルミジンアセチルトランスフェラーゼのX線結晶構造解析(蛋白質-構造,第48回日本生物物理学会年会)
• Control of Protein Crystal Nucleation and Growth Using Stirring Solution
• Ni_Co_x-C Nanogranular Thin Films Prepared by a Co-S puttering Method:Improvement in Magnetic Properties by Optimizing the Alloy Ratio : Magnetism
• Thin Films of Carbon Nanocapsules and Onion-Like Graphitic Particles Prepared by the Cosputtering Method
• In Situ Monitoring of Photodynamic Inactivation of the Membrane Functions of Bacteria Using Electrochemical Sensors
• Application of an Oxygen Electrode to Evaluate Superoxide Anion-scavenging Ability
• Precise Size Determination of Amphotericin B and Nystatin Channels Formed in Erythrocyte and Liposomal Membranes Based on Osmotic Protection Experiments
• 2P066 1E1450 巨大分子量タンパク質の構造安定性(蛋白質-物性(安定性,折れたたみなど),第48回日本生物物理学会年会)
• 1P-020 Mechanism of Interfacial Activation of a Family I.3 Lipase with Two Lid Structures Revealed by X-ray Crystallography and Molecular Dynamics Simulation(Protein:Structure & Function, The 47th Annual Meeting of the Biophysical Society of Japan)
• Promotion of Crystal Nucleation of Protein by Semi-Solid Agarose Gel
• Evaluation and improvement of a technique to manipulate protein crystals in solution
• Partial Ferromagnetism in the Hubbard Chain with Next-Nearest-Neighbor Hopping (Condensed Matter: Electronic Structure, Electrical, Magnetic and Optical Properties)
• 387-nm Generation in Gd_xY_Ca_4O(BO_3)_3 Crystal and Its Utilization for 193-nm Light Source
• A Palm-Size Ultraviolet Laser Using a Combination of a Monolithic Wavelength Converter and an Optical Fiber
• Drude Weight of the Two-Dimensional Hubbard Model : Reexamination of Finite-Size Effect in Exact Diagonalization Study(Condensed matter: electronic structure and electrical, magnetic, and optical properties)
• Effect of Evaporation on Protein Crystals Grown in Semi-Solid Agarose Hydrogel
• Generation of Tunable Near-UV Laser Radiation by Type-I Second-Harmonic Generation in a New Crystal, K2Al2B2O7 (KABO)
• New Practical Technique for Protein Crystallization with Floating and Stirring Methods
• Bulk Crystal Growth of Stilbazolium Derivatives for Terahertz Waves Generation
• Synthesis of GaN Crystal Using Gallium Hydride
• Two-Liquid Hanging-Drop Vapor-Diffusion Technique of Protein Crystallization
• A Phalaenopsis variety with floral organs showing C class homeotic transformation and its revertant may enable Phalaenopsis as a potential molecular genetic material
• Femtosecond Laser Processing of Agarose Gel Surrounding Protein Crystals for Development of an Automated Crystal Capturing System
• Protein Crystallization in Agarose Gel with High Strength: Developing an Automated System for Protein Crystallographic Processes
• Crystal structure of peroxiredoxin from Aeropyrum pernix K1 complexed with its substrate, hydrogen peroxide
• 2P213 Processive movement of minus end-directed microtubule motor protein Pkl1(37. Molecular motor (II),Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)
• Nonpolar a-Plane AlGaN/GaN Heterostructure Field-Effect Transistors Grown on Freestanding GaN Substrate
• Spatio-Temporal Control of Bacterial-Suspension Luminescence Using a PDMS Cell
• Metal-Insulator Transition of the Hubbard Chain with Next-Nearest-Neighbor Hopping : Partially Ferromagnetic Metal toward Mott Insulator (Condensed Matter : Electronic Structure, Electrical, Magnetic and Optical Properties)
• A Manipulating Tool for Protein Microcrystals in Solution Using Adhesive Materials
• Evaluation and Improvement of a Technique to Manipulate Protein Crystals in Solution
• A Phalaenopsis variety with floral organs showing C class homeotic transformation and its revertant may enable Phalaenopsis as a potential molecular genetic material
• Gruneisen's Approach to Magnetovolume Effect of Itinerant Electron Ferromagnets
• Itinerant Electron Metamagnetism in $\eta$-Carbide-Type Compound Co3Mo3C
• Magnetization Process of Narrow-Gap Semiconductor FeSb2
• Examination of Effects of H2 Concentration in Reactant Gas on GaN Growth by Gallium Hydride Vapor Phase Epitaxy
• Growth of Thick GaN Films with High Growth Rate Using Sublimation Method under High Pressure
• Protein Crystal Growth Using Laser-Processed Seed Crystals
• Structural insight into the stereoselective production of PGF_ by Old Yellow Enzyme from Trypanosoma cruzi
• Phase-Matching Properties at around 190nm of Various Borate Crystals
• Protein Crystallization by Combining Laser Irradiation and Solution-Stirring Techniques
• Polymorphs of Rubrene Crystal Grown from Solution
• Solution Growth of Rubrene Single Crystals Using Various Organic Solvents
• Drastic Decrease in Dislocations during Liquid Phase Epitaxy Growth of GaN Single Crystals Using Na flux Method without Any Artificial Processes
• Synthesis of AlN Grains and Liquid-Phase-Epitaxy (LPE) Growth of AlN Films Using Sn-Ca Mixed Flux
• Temperature-Screening System for Determining Protein Crystallization Conditions
• Structural Analysis of Carbon-Added Na--Ga Melts in Na Flux GaN Growth by First-Principles Calculation
• Femtosecond Laser Processing of Protein Crystals in Crystallization Drop
• 387-nm Generation in GdxY1-xCa4O(BO3)3 Crystal and Its Utilization for 193-nm Light Source
• Structural Analysis of Carbon-Added Na-Ga Melts in Na Flux GaN Growth by First-Principles Calculation (Special Issue : Recent Advances in Nitride Semiconductors)
• Semiautomatic Protein Crystallization System Featuring Crystallization Solution Preparation Function
• Al Doping of CsLiB_6O_ for High Resistance to Ultraviolet-Induced Degradation

もっと見る 閉じる

スポンサーリンク