Dataset Information

Fast Fluoroalkylation of Proteins Uncovers Structure and Dynamics of Biological Macromolecules

ABSTRACT: Stable covalent labeling of proteins in combination with mass spectrometry has been established as a complementary technique to classical structural methods, such as X Ray, NMR, or Cryo-EM used for protein structure determination. Although the current stable covalent labeling techniques enable to monitor protein solvent accessible areas with sufficient spatial resolution, there is still high demand for alternative, less complicated and inexpensive approaches. Here, we introduce a new covalent labeling method based on Fast Fluoroalkylation of Proteins (FFAP). The FFAP uses fluoroalkyl radicals formed by reductive decomposition of Togni reagents with ascorbic acid for labeling of proteins on a time scale of seconds. The feasibility of FFAP for effective labeling of proteins was demonstrated by monitoring differential amino acids modi-fication of native horse heart apomyoglobin/holomyoglobin and human haptoglobin-hemoglobin complex. The obtained data confirmed the Togni reagent-mediated FFAP as an advantageous alternative method for covalent labeling in applications, such as protein fingerprinting, epitope mapping and surface mapping of proteins (and their complexes) in general.

INSTRUMENT(S): Bruker Daltonics solarix series

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Zdenek Kukacka

LAB HEAD: Petr Novak

PROVIDER: PXD027310 | Pride | 2021-12-30

REPOSITORIES: pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 50

Publications

Fast Fluoroalkylation of Proteins Uncovers the Structure and Dynamics of Biological Macromolecules.

Fojtík Lukáš L Fiala Jan J Pompach Petr P Chmelík Josef J Matoušek Václav V Beier Petr P Kukačka Zdeněk Z Novák Petr P

Journal of the American Chemical Society 20211130 49

Covalent labeling of proteins in combination with mass spectrometry has been established as a complementary technique to classical structural methods, such as X-ray, NMR, or cryogenic electron microscopy (Cryo-EM), used for protein structure determination. Although the current covalent labeling techniques enable the protein solvent accessible areas with sufficient spatial resolution to be monitored, there is still high demand for alternative, less complicated, and inexpensive approaches. Here, w ...[more]

PMID: 34846870

Similar Datasets

Project description:Background: Progressive neurological dysfunction is a key aspect of human aging. Because of underlying differences in the aging of mice and humans, useful mouse models have been difficult to obtain and study. We have used gene-expression analysis and polymorphism screening to study molecular senescence of the retina and hippocampus in two rare inbred mouse models of accelerated neurological senescence (SAMP8 and SAMP10) that closely mimic human neurological aging, and in a related normal strain (SAMR1) and an unrelated normal strain (C57BL/6J). Results: The majority of age-related gene expression changes were strain-specific, with only a few common pathways found for normal and accelerated neurological aging. Polymorphism screening led to the identification of mutations that could have a direct impact on important disease processes, including a mutation in a fibroblast growth factor gene, Fgf1, and a mutation in and ectopic expression of the gene for the chemokine CCL19, which is involved in the inflammatory response. Conclusions: We show that combining the study of inbred mouse strains with interesting traits and gene-expression profiling can lead to the discovery of genes important for complex phenotypes. Furthermore, full-genome polymorphism detection, sequencing and gene-expression profiling of inbred mouse strains with interesting phenotypic differences may provide unique insights into the molecular genetics of late-manifesting complex diseases. Experiment Overall Design: Gene-expression profiling was performed on 3 month-old (young), 16 month-old (old) S8, S10 and SR1 mice. Two independent samples for each time point were used in gene-expression profiling for each strain. Because of greater replicate variability, three samples were used for hippocampus of 16-month SAM mice. A 10.0 ug sample of total RNA was used to generate labeled cRNA for each sample according to recommended protocols (Affymetrix). RNA from multiple animals was not pooled, except in the case of retina, where the retinas of two mice were combined to generate sufficient total RNA.

			Action	DRS
	APO-IM-ACmgf.mgf	Mgf
	APO-IM-OHmgf.mgf	Mgf
	APO-T-ACmgf.mgf	Mgf
	APO-T-OHmgf.mgf	Mgf
	F006428.dat	Other