Proteomics

Dataset Information

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TULIP2: an improved method for the identification of ubiquitin E3-specific targets


ABSTRACT: Protein modification by Ubiquitin or Ubiquitin-like modifiers is mediated by an enzyme cascade composed of E1s, E2s and E3s enzymes. E1, or ubiquitin-activating enzymes, perform the ubiquitin activation. Next, ubiquitin is transferred to ubiquitin-conjugating enzymes or E2s. Finally, ubiquitin ligases or E3s catalyze the transfer of ubiquitin to the acceptor proteins. E3 enzymes are responsible of determining the substrate specificity. Determining which E3 enzyme maps to which substrate is a major challenge that is greatly facilitated by the TULIP2 methodology. TULIP2 methodology is fast, precise and cost-effective. Compared to previous TULIP methodology protocol, TULIP2 methodology achieves a more than 50-fold improvement in the purification yield and two orders of magnitude improvement in the signal-to-background ratio after label free quantification by mass spectrometry analysis. The method includes the generation of TULIP2 cell lines, subsequent purification of TULIP2 conjugates, preparation and analysis of samples by mass spectrometry

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo sapiens  

TISSUE(S): Tissue Not Applicable To Dataset

DISEASE(S): Not Available

SUBMITTER: Román González-Prieto  

LAB HEAD: Román González-Prieto

PROVIDER: PXD015437 | Pride | 2019-11-15

REPOSITORIES: pride

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Publications

TULIP2: An Improved Method for the Identification of Ubiquitin E3-Specific Targets.

Salas-Lloret Daniel D   Agabitini Giulia G   González-Prieto Román R  

Frontiers in chemistry 20191126


Protein modification by Ubiquitin or Ubiquitin-like modifiers is mediated by an enzyme cascade composed of E1, E2, and E3 enzymes. E1s, or ubiquitin-activating enzymes, perform ubiquitin activation. Next, ubiquitin is transferred to ubiquitin-conjugating enzymes or E2s. Finally, ubiquitin ligases or E3s catalyze the transfer of ubiquitin to the acceptor proteins. E3 enzymes are responsible for determining the substrate specificity. Determining which E3 enzyme maps to which substrate is a major c  ...[more]

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