NDLI: Systematic determinations of SUMOylation activation intermediates and dynamics by a sensitive and quantitative FRET assay

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Hypothesis: butyrate is not an HDAC inhibitor, but a product inhibitor of deacetylation

Proteoglycan sequence

Genome-scale genetic manipulation methods for exploring bacterial molecular biology

Endothelial cell's biomechanical properties are regulated by invasive cancer cells

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The lipid composition of isolated cytoplasmic lipid droplets from a human cancer cell line, BE(2)M17

Probing adenylation: using a fluorescently labelled ATP probe to directly label and immunoprecipitate VopS substrates

Positive regulatory dynamics by a small noncoding RNA: speeding up responses under temperature stress

Naïve Bayes QSDR classification based on spiral-graph Shannon entropies for protein biomarkers in human colon cancer

Systematic determinations of SUMOylation activation intermediates and dynamics by a sensitive and quantitative FRET assay

Proteomic analysis of a drosophila IBMPFD model reveals potential pathogenic mechanisms

Fabrication of nanostructures through molecular self-assembly of small amphiphilic glyco-dehydropeptides

Insertion of T4-lysozyme (T4L) can be a useful tool for studying olfactory-related GPCRs

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E. coli LoiP (YggG), a metalloprotease hydrolyzing Phe–Phe bonds

Metabolomic analysis of the plasma of patients with high-altitude pulmonary edema (HAPE) using ¹H NMR

Integrated investigation of lipidome and related signaling pathways uncovers molecular mechanisms of tetramethylpyrazine and butylidenephthalide protecting endothelial cells under oxidative stress

Backbone conformational preferences of an intrinsically disordered protein in solution

Proteolysis of beta-galactosidase following SigmaB activation in Bacillus subtilis

Engineering a large protein by combined rational and random approaches: stabilizing the Clostridium thermocellum cellobiose phosphorylase

Rewiring and dosing of systems modules as a design approach for synthetic mammalian signaling networks

An orthogonal comparison of the proteome of human embryonic stem cells with that of human induced pluripotent stem cells of different genetic background

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Systematic determinations of SUMOylation activation intermediates and dynamics by a sensitive and quantitative FRET assay

Content Provider	Royal Society of Chemistry (RSC)
Author	Liao, Jiayu Song, Yang
Copyright Year	2012
Abstract	Ubiquitination and SUMOylation are multi-step cascade reactions, in which small protein modifiers are activated by E1 activating enzyme, transferred to E2 conjugating enzyme, and conjugated to substrates mediated by the E3 ligase in vivo. The structural and biochemical bases for the cascade reactions have been elucidated by several studies. However, the reaction intermediates and dynamics of these peptide modifiers among the enzymes have not been completely elucidated. Here we report detailed investigations of SUMOylation dynamics and interaction switches of SUMO1 among its ligases using FRET technology. These studies show that, while SUMO1 and the E1 subunit Aos1 or Uba2 have no intrinsic affinity for each other, the adenylation of SUMO1 carried out by Aos1 requires the presence of Uba2, and subsequently conformational changes trigger the interaction of SUMO1 and Uba2 for a thioester bond formation. The reaction intermediates among SUMO1 and its ligases are indirectly revealed by FRET signals generated by each pair. Furthermore, the transfer of SUMO1 from Uba2 to E2 enzyme, Ubc9, depends on the formation of a thioester bond between SUMO1 and Ubc9, and requires non-covalent interaction between Ubc9 and Uba2, but not between Ubc9 and SUMO1. These interaction switches provide the physical and biochemical bases for the SUMO activation and a transfer cascade required for SUMO activation.
Starting Page	1723
Ending Page	1729
Page Count	7
File Format	HTM / HTML PDF
ISSN	1742206X
Volume Number	8
Issue Number	6
Journal	Molecular BioSystems
DOI	10.1039/c2mb05465e
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	SUMO protein Ligase Förster resonance energy transfer Biological target Enzyme Peptide bond Thioester Covalent bond Small ubiquitin-related modifier 1 Protein
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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