Macromolecular Cryocrystallography: Cooling, Mounting, Storage and Transportation of Crystals

Parkin, Sean; Hope, H.

doi:10.1107/s0021889898005305

Cited by 87 publications

(71 citation statements)

References 13 publications

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“…Lysozyme samples were mounted on a magnetic goniometer head by rapid transfer from liquid nitrogen into the cold nitrogen stream using a CryoTong (Hampton Research), following standard procedures (Hope, 1990;Rodgers, 1994;Young et al, 1994;Kurinov & Harrison, 1995;Garman & Schneider, 1997;Parkin & Hope, 1998;Garman, 1999). Flow temperatures were 150 K. Measurements of liquid-nitrogen¯ash-cooled crystals at temperatures between 100 and 200 K using a laboratory X-ray source showed that the diffraction properties did not degrade during several hours at 150 K. At this temperature Young's modulus and other crystal mechanical properties are nearly temperature independent (Morozov & Gevorkian, 1985) and appreciable relaxation of amorphous ice to crystalline ice does not occur on the 30±120 min timescale of our experiments (Miyazaki et al, 2000).…”

Section: Methodsmentioning

confidence: 99%

Flash-cooling and annealing of protein crystals

Kriminski

Caylor

Nonato

et al. 2002

Acta Crystallogr D Biol Crystallogr

107

162

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Flash-cooling and annealing of macromolecular crystals have been investigated using in situ X-ray imaging, diffraction-peak lineshape measurements and conventional crystallographic diffraction. The dominant mechanisms by which¯ash-cooling creates disorder are suggested and a ®xed-temperature annealing protocol for reducing this disorder is demonstrated that should be more reliable and¯exible than existing protocols. Flash-cooling tetragonal lysozyme crystals degrades diffraction resolution and broadens the distributions of lattice orientations (mosaicity) and lattice spacings. The diffraction resolution strongly correlates with the width of the latticespacing distribution. Annealing at ®xed temperatures of 253 and 233 K consistently reduces the lattice-spacing spread and improves the resolution for annealing times up to $30 s. X-ray images show that this improvement arises from the formation of well ordered domains with characteristic sizes >10 mm and narrower mosaicities than the crystal as a whole. Flash-cooled triclinic crystals of lysozyme, which have a smaller water content than the tetragonal form, diffract to higher resolution with smaller mosaicities and exhibit pronounced ordered domain structure even before annealing. It is suggested that differential thermal expansion of the protein lattice and solvent may be the primary cause of¯ash-cooling-induced disorder. Mechanisms by which annealing at T << 273 K reduce this disorder are discussed.

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Section: Methodsmentioning

confidence: 99%

Flash-cooling and annealing of protein crystals

Kriminski

Caylor

Nonato

et al. 2002

Acta Crystallogr D Biol Crystallogr

107

162

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“…This consists of a tiny nylon loop mounted on a small stainless steel tube inserted into the lid of a cryovial. The cryoloop, was first applied to the flash freezing of protein solutions for analysis in crystallography (Parkin and Hope, 1998), and was later used to cryopreserve mammalian embryos and oocytes (Lane et al, 2001), resulting in births from vitrified blastocysts in the case of humans (Makkaida et al, 2001) and monkeys (Makkaida et al, 2003). Nevertheless, despite its successes, the cryoloop is a sensitive and fragile system that may increase the risk of accidental warming (Kuwayama, 2007).…”

Section: New Trends In Vitrification Of Oocytes and Embryosmentioning

confidence: 99%

Oocyte and embryo cryopreservation – state of art and recent developments in domestic animals

Gajda¹,

Smorąg²

2009

J. Anim. Feed Sci.

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During the past few decades, significant progress in cryopreservation of mammalian oocytes and embryos has been observed. Transfer of cryopreserved embryos or oocytes resulted in live offspring in at least 25 species. So far, two major methods have been used for oocyte and embryo cryopreservation: conventional slow-rate freezing and vitrification. This review summarizes the progress in cryopreservation of mammalian oocytes and embryos that has been achieved by modifying oocyte/embryo susceptibility to cryopreservation or vitrification technology through such techniques as solid-surface vitrification, cryoloop, microdrop, cryotop, electron microscopy grids, nylon mesh, open pulled straw method or removal of lipids, addition of antifreeze protein or cytoskeletonstabilizing agents, cholesterol or liposomes, centrifugation prior to cryopreservation or application of high hydrostatic pressure. In conclusion, the vitrification method opened new perspectives in cryopreservation of embryos and oocytes, both for in vitro fertilized and somatic nuclear transfer. Authors believe that new cryopreservation procedures which modify the susceptibility of oocytes and embryos to cryopreservation will gain importance as a major tool in mammalian gamete and embryo cryopreservation.

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“…After incubation at 277 K for 24-48 h, the soluble portion of the complexes were concentrated and purified by size exclusion and subsequently by anionexchange chromatography. The three pHLA-B*3901 complexes were finally concentrated to 4-8 mg/ml (20 (28). X-ray diffraction data were collected at beamline BL17U on Shanghai Synchrotron Radiation Facility.…”

Section: Expression Purification Crystallization and Data Collectionmentioning

confidence: 99%

Nα-Terminal Acetylation for T Cell Recognition: Molecular Basis of MHC Class I–Restricted Nα-Acetylpeptide Presentation

Sun

et al. 2014

The Journal of Immunology

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As one of the most common posttranslational modifications (PTMs) of eukaryotic proteins, Nα-terminal acetylation (Nt-acetylation) generates a class of Nα-acetylpeptides that are known to be presented by MHC class I at the cell surface. Although such PTM plays a pivotal role in adjusting proteolysis, the molecular basis for the presentation and T cell recognition of Nα-acetylpeptides remains largely unknown. In this study, we determined a high-resolution crystallographic structure of HLA (HLA)-B*3901 complexed with an Nα-acetylpeptide derived from natural cellular processing, also in comparison with the unmodified-peptide complex. Unlike the α-amino–free P1 residues of unmodified peptide, of which the α-amino group inserts into pocket A of the Ag-binding groove, the Nα-linked acetyl of the acetylated P1-Ser protrudes out of the groove for T cell recognition. Moreover, the Nt-acetylation not only alters the conformation of the peptide but also switches the residues in the α1-helix of HLA-B*3901, which may impact the T cell engagement. The thermostability measurements of complexes between Nα-acetylpeptides and a series of MHC class I molecules derived from different species reveal reduced stability. Our findings provide the insight into the mode of Nα-acetylpeptide–specific presentation by classical MHC class I molecules and shed light on the potential of acetylepitope-based immune intervene and vaccine development.

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Macromolecular Cryocrystallography: Cooling, Mounting, Storage and Transportation of Crystals

Cited by 87 publications

References 13 publications

Flash-cooling and annealing of protein crystals

Flash-cooling and annealing of protein crystals

Oocyte and embryo cryopreservation – state of art and recent developments in domestic animals

Nα-Terminal Acetylation for T Cell Recognition: Molecular Basis of MHC Class I–Restricted Nα-Acetylpeptide Presentation

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