Sumoylation Ubiquitination
Sumoylation Ubiquitination - Related Videos
06:00
SUMOylation Assay: An In Vitro Technique to Detect the SUMOylation Status of Substrate Proteins by Immunoblotting
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2025
This video demonstrates the in vitro method for SUMOylation of substrate proteins using a sequential enzyme cascade. Further, the SUMOylated status of the protein is identified using the electrophoresis and immunoblotting technique.
12:28
Protein Purification Technique that Allows Detection of Sumoylation and Ubiquitination of Budding Yeast Kinetochore Proteins Ndc10 and Ndc80
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Cited by 11 •
2015
This manuscript describes the detection of sumoylation and ubiquitination of kinetochore proteins, Ndc10 and Ndc80, in the budding yeast Saccharomyces cerevisiae.
09:40
In Vivo Detection and Analysis of Rb Protein SUMOylation in Human Cells
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Cited by 2 •
2017
Small ubiquitin-related modifier (SUMO) family proteins are conjugated to the lysine residues of target proteins to regulate various cellular processes. This paper describes a protocol for the detection of retinoblastoma (Rb) protein SUMOylation under endogenous and exogenous conditions in human cells.
09:00
Detection of Protein Ubiquitination
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Cited by 96 •
2009
Ubiquitination is a key posttranslational modification carried out by a set of three enzymes. Mutations of genes involved in this modification are associated with many different human diseases. Here, we describe protocols to detect protein ubiquitination in cultured cells in vivo and test tubes in vitro.
10:43
siRNA Screening to Identify Ubiquitin and Ubiquitin-like System Regulators of Biological Pathways in Cultured Mammalian Cells
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Cited by 4 •
2014
Here, we describe a methodology to perform a targeted siRNA “ubiquitome” screen to identify novel ubiquitin and ubiquitin-like regulators of the HIF1A-mediated cellular response to hypoxia. This can be adapted to any biological pathway where a robust read out of reporter activity is available.
FAQs
The modifier attached to a protein strongly influences the resulting cellular response. Ubiquitination can direct a target toward proteasomal degradation, linking the modification to protein turnover and quality control. Sumoylation more commonly changes nuclear transport, transcription, DNA repair, stress responses, or molecular interactions without necessarily destroying the target. This distinction helps researchers interpret modification patterns within signaling pathways.
E1-activating enzymes initiate the modification cascade, E2-conjugating enzymes carry the activated modifier, and E3-ligating enzymes promote its attachment to a target protein. Together, these enzyme classes provide an organized route for modifying selected substrates, often at lysine residues. Examining this cascade helps investigators determine where regulation occurs and how altered enzyme activity may affect protein stability or function.
SUMO and ubiquitin are not interchangeable regulatory signals. Attaching SUMO commonly influences a protein’s localization, transcriptional activity, DNA-repair participation, stress response, or interactions, whereas ubiquitination may contribute to proteasomal removal. Consequently, identifying which modifier is present provides essential context for interpreting a protein’s behavior, particularly when studying signaling, protein quality control, or changes in cellular activity.
Detection-focused techniques can be used to determine whether a target protein carries SUMO or ubiquitin, while manipulation-focused techniques can alter the modification process or its enzyme cascade. Researchers can then relate these changes to protein stability, localization, activity, or interactions. This combined strategy connects a molecular modification with its cellular consequence rather than treating its presence as an isolated observation.
Protein abundance alone may not reveal whether a molecule has changed location, activity, interaction partners, or susceptibility to degradation. Studying sumoylation and ubiquitination adds information about regulatory state and protein quality control. Researchers therefore examine these modifications when characterizing signaling pathways, stress responses, DNA-repair processes, or mechanisms that alter cellular protein behavior without simply changing total protein levels.
Both modification systems provide a framework for examining disease mechanisms that involve signaling, protein regulation, and quality control. In cancer research, investigators can study how altered modification-dependent regulation affects cellular pathways. In neurodegeneration research, the same approaches help examine disrupted protein handling and stability. Detecting or manipulating these processes can therefore connect molecular changes with disease-associated cellular dysfunction.