Learning about Biomolecular Solvation from Water in Protein Crystals

RuBisCO (D-ribulose-l,5-biphosphate carboxylase/oxygenase) is the most aboundant enzyme in the plant cell and it catalyses the key carboxylation reaction of photosynthetic carbon fixation, but also the competing oxygenase reaction of photorespiation. In vitro crystallized RuBisCO has been studied earlier but this investigation concerns in vivo existance of RuBisCO crystals in anthers and leaves ofsugarbeets. For the identification of in vivo protein crystals it is important to be able to determinethe unit cell of cytochemically identified crystals in the same image. In order to obtain the best combination of optimal contrast and resolution we have studied different staining and electron accelerating voltages. It is known that embedding and sectioning can cause deformation and obscure the unit cell parameters.

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Dynamical Scattering in Electron Diffraction of wet Protein Crystals

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s1431927600000167 ◽

1980 ◽

Vol 38 ◽

pp. 42-43

Author(s):

B. B. Chang ◽

D. F. Parsons

Keyword(s):

Electron Diffraction ◽

Scattering Theory ◽

Protein Crystals ◽

Electron Diffraction Data ◽

Specimen Thickness ◽

Major Question ◽

X Ray ◽

Scattering Effects ◽

Diffraction Patterns ◽

Acceleration Voltage

The significance of dynamical scattering effects remains the major question in the structural analysis by electron diffraction of protein crystals preserved in the hydrated state. In the few cases (single layers of purple membrane and 400-600 Å thick catalase crystals examined at 100 kV acceleration voltage) where electron-diffraction patterns were used quantitatively, dynamical scattering effects were considered unimportant on the basis of a comparison with x-ray intensities. The kinematical treatment is usually justified by the thinness of the crystal. A theoretical investigation by Ho et al. using Cowley-Moodie multislice formulation of dynamical scattering theory and cytochrome b5as the test object2 suggests that kinematical analysis of electron diffraction data with 100-keV electrons would not likely be valid for specimen thickness of 300 Å or more. We have chosen to work with electron diffraction patterns obtained from actual wet protein crystals (rat hemoglobin crystals of thickness range 1000 to 2500 Å) at 200 and 1000 kV and to analyze these for dynamical effects.

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Langmuir-Blodgett Nanotemplate and Radiation Resistance in Protein Crystals: State of the Art

Critical Reviews in Eukaryotic Gene Expression ◽

10.1615/critreveukargeneexpr.v22.i3.50 ◽

2012 ◽

Vol 22 (3) ◽

pp. 219-232 ◽

Cited By ~ 3

Author(s):

Luca Belmonte ◽

Eugenia Pechkova ◽

Shailesh Tripathi ◽

Dora Scudieri ◽

Claudio Nicolini

Keyword(s):

Radiation Resistance ◽

State Of The Art ◽

Protein Crystals ◽

Langmuir Blodgett

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Analysis of Element Composition of DNA-Protein Crystals In Vitro

Moscow University Biological Sciences Bulletin ◽

10.3103/s0096392519040102 ◽

2019 ◽

Vol 74 (4) ◽

pp. 240-245

Author(s):

A. V. Moiseenko ◽

N. G. Loiko ◽

O. V. Chertkov ◽

A. V. Feofanov ◽

Yu. F. Krupyanskii ◽

...

Keyword(s):

Element Composition ◽

Protein Crystals

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Technology of high quality protein crystals’ obtaining aboard the ISS at execution of joint Japanese–Russian experiments

Space engineering and technology ◽

10.33950/spacetech-2308-7625-2020-2-5-25 ◽

2020 ◽

pp. 5-25

Author(s):

Koji INAKA ◽

Saori ICHIMIZU ◽

Izumi YOSHIZAKI ◽

Kiyohito KIHIRA ◽

Elena G. LAVRENKO ◽

...

Keyword(s):

Drug Design ◽

International Space Station ◽

Space Station ◽

Protein Crystals ◽

Diffusion Method ◽

X Ray Diffraction ◽

High Quality ◽

International Space ◽

X Ray ◽

High Quality Protein

A series of space experiments aboard the International Space Station (ISS) associated with high-quality Protein Crystal Growth (PCG) in microgravity conditions can be considered as a unique and one of the best examples of fruitful collaboration between Japanese and Russian scientists and engineers in space, which includes also other ISS International Partners. X-ray diffraction is still the most powerful tool to determine the protein three dimensional structure necessary for Structure based drug design (SBDD). The major purpose of the experiment is to grow high quality protein crystals in microgravity for X-ray diffraction on Earth. Within one and a half decade, Japan and Russia have established an efficient process over PCG in space to support latest developments over drug design and structural biology. One of the keys for success of the experiment lies in how precisely pre-launch preparations are made. Japanese party provides flight equipment for crystallization and ensures the required environment to support the experiment aboard of the ISS’s Kibo module, and also mainly takes part of the experiment ground support such as protein sample characterization, purification, crystallization screening, and solution optimization for microgravity experiment. Russian party is responsible for integration of the flight items equipped with proteins and precipitants on board Russian transportation space vehicles (Soyuz or Progress), for delivery them at the ISS, transfer to Kibo module, and returning the experiments’ results back on Earth aboard Soyuz manned capsule. Due to close cooperation of the parties and solid organizational structure, samples can be launched at the ISS every half a year if the ground preparation goes smoothly. The samples are crystallized using counter diffusion method at 20 degree C for 1–2.5 months. After samples return, the crystals are carefully taken out from the capillary, and frozen for X-ray diffraction at SPring8 facility in Japan. Extensive support of researchers from both countries is also a part of this process. The paper analyses details of the PCG experiment scheme, unique and reliable technology of its execution, and contains examples of the application. Key words: International Space Station, Protein crystals, Microgravity, International collaboration.

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