Utilization of a very small anomalous diffraction signal provided by sulfur atoms of the cysteine and methionine residues or by phosphorus atoms of nucleic acids, intrinsically present in all proteins or RNA/DNA can be successfully used to solve crystal structures of proteins. This approach, in part pioneered by us, is currently gaining increasing popularity and becomes a routine technique in the practice of macromolecular crystallography. The diffraction data to extremely high resolution in the range of 0.6-0.8 angstrom were measured using synchrotron radiation from crystals of oligonucleotides and oligopeptides, and the refined models revealeed the unprecedented structural details of the investigated molecules including electron densities at the sigma bonds between atoms. The observed very fine features in this structures will serve to improve the stereochemical libraries used for routine refinement of protein crystal structures at lower resolution. We participated in several collaborative structural projects on various biologically important proteins, such as fluorescenty proteins used as biomarkers, human mitochindrial helicase, rhizobium fucosyltransferase and plant S-adenosyl-homocysteine hydrolase. We performed the accurate evaluation of the effects of X-ray radiation damage incurred in macromolecular crystals and established the limit of acceptable absorption dose at 7 MGy.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC010378-16
Application #
9153556
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
16
Fiscal Year
2015
Total Cost
Indirect Cost
Name
Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Ruszkowski, Milosz; Sekula, Bartosz; Ruszkowska, Agnieszka et al. (2018) Chloroplastic Serine Hydroxymethyltransferase From Medicago truncatula: A Structural Characterization. Front Plant Sci 9:584
Luo, Zhipu; Dauter, Zbigniew (2018) The crystal structure of Z-DNA with untypically coordinated Ca2+ ions. J Biol Inorg Chem 23:253-259
Wlodawer, Alexander; Dauter, Zbigniew; Porebski, Przemyslaw J et al. (2018) Detect, correct, retract: How to manage incorrect structural models. FEBS J 285:444-466
Dauter, Zbigniew; Jaskolski, Mariusz (2018) On the helical arrangements of protein molecules. Protein Sci 27:643-652
Dauter, Zbigniew; Wlodawer, Alexander (2018) Unexpected Specificity of a Trypsin-like Enzyme. Structure 26:530-531
Sekula, Bartosz; Ruszkowski, Milosz; Dauter, Zbigniew (2018) Structural Analysis of Phosphoserine Aminotransferase (Isoform 1) From Arabidopsis thaliana- the Enzyme Involved in the Phosphorylated Pathway of Serine Biosynthesis. Front Plant Sci 9:876
Sekula, Bartosz; Dauter, Zbigniew (2018) Crystal structure of thermospermine synthase from Medicago truncatula and substrate discriminatory features of plant aminopropyltransferases. Biochem J 475:787-802
Pyra, Anna; Brzozowska, Ewa; Pawlik, Krzysztof et al. (2017) Tail tubular protein A: a dual-function tail protein of Klebsiella pneumoniae bacteriophage KP32. Sci Rep 7:2223
Luo, Zhipu; Dauter, Zbigniew (2017) Embarras de richesses - It is not good to be too anomalous: Accurate structure of selenourea, a chiral crystal of planar molecules. PLoS One 12:e0171740
Dauter, Miroslawa; Dauter, Zbigniew (2017) Many Ways to Derivatize Macromolecules and Their Crystals for Phasing. Methods Mol Biol 1607:349-356

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