Identification of SUMO targets through in vitro expression cloning

Christian B. Gocke, Hongtao Yu

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Citation (Scopus)

Abstract

Covalent modification of proteins by small ubiquitin-like modifier (SUMO) regulates diverse cellular processes. While many SUMO substrates are identified through individual efforts, affinity-based approaches followed by mass spectrometry are also used to identify in vivo SUMO substrates in yeast and in mammals. Because of low steady-state levels of sumoylation and biases towards abundant targets, identifying sumoylated proteins in vivo can be challenging. The in vitro expression cloning (IVEC) method for SUMO target identification circumvents these challenges and complements the affinity-based approaches. IVEC allows for immediate validation and analysis of substrates through in vitro reconstitution. Furthermore, this method can be easily adapted to identify substrates of specific SUMO ligases.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
Pages51-61
Number of pages11
Volume497
DOIs
StatePublished - 2009

Publication series

NameMethods in Molecular Biology
Volume497
ISSN (Print)10643745

Fingerprint

Ubiquitin
Organism Cloning
Ubiquitins
Sumoylation
Ligases
Mammals
Mass Spectrometry
Yeasts
In Vitro Techniques
Proteins

Keywords

  • In vitro expression cloning
  • SUMO
  • SUMO ligase
  • Sumoylation
  • Ubc9

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Gocke, C. B., & Yu, H. (2009). Identification of SUMO targets through in vitro expression cloning. In Methods in Molecular Biology (Vol. 497, pp. 51-61). (Methods in Molecular Biology; Vol. 497). https://doi.org/10.1007/978-1-59745-566-4_4

Identification of SUMO targets through in vitro expression cloning. / Gocke, Christian B.; Yu, Hongtao.

Methods in Molecular Biology. Vol. 497 2009. p. 51-61 (Methods in Molecular Biology; Vol. 497).

Research output: Chapter in Book/Report/Conference proceedingChapter

Gocke, CB & Yu, H 2009, Identification of SUMO targets through in vitro expression cloning. in Methods in Molecular Biology. vol. 497, Methods in Molecular Biology, vol. 497, pp. 51-61. https://doi.org/10.1007/978-1-59745-566-4_4
Gocke CB, Yu H. Identification of SUMO targets through in vitro expression cloning. In Methods in Molecular Biology. Vol. 497. 2009. p. 51-61. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-59745-566-4_4
Gocke, Christian B. ; Yu, Hongtao. / Identification of SUMO targets through in vitro expression cloning. Methods in Molecular Biology. Vol. 497 2009. pp. 51-61 (Methods in Molecular Biology).
@inbook{5980b969912b4934958adb7dd9c98c62,
title = "Identification of SUMO targets through in vitro expression cloning",
abstract = "Covalent modification of proteins by small ubiquitin-like modifier (SUMO) regulates diverse cellular processes. While many SUMO substrates are identified through individual efforts, affinity-based approaches followed by mass spectrometry are also used to identify in vivo SUMO substrates in yeast and in mammals. Because of low steady-state levels of sumoylation and biases towards abundant targets, identifying sumoylated proteins in vivo can be challenging. The in vitro expression cloning (IVEC) method for SUMO target identification circumvents these challenges and complements the affinity-based approaches. IVEC allows for immediate validation and analysis of substrates through in vitro reconstitution. Furthermore, this method can be easily adapted to identify substrates of specific SUMO ligases.",
keywords = "In vitro expression cloning, SUMO, SUMO ligase, Sumoylation, Ubc9",
author = "Gocke, {Christian B.} and Hongtao Yu",
year = "2009",
doi = "10.1007/978-1-59745-566-4_4",
language = "English (US)",
isbn = "9781934115800",
volume = "497",
series = "Methods in Molecular Biology",
pages = "51--61",
booktitle = "Methods in Molecular Biology",

}

TY - CHAP

T1 - Identification of SUMO targets through in vitro expression cloning

AU - Gocke, Christian B.

AU - Yu, Hongtao

PY - 2009

Y1 - 2009

N2 - Covalent modification of proteins by small ubiquitin-like modifier (SUMO) regulates diverse cellular processes. While many SUMO substrates are identified through individual efforts, affinity-based approaches followed by mass spectrometry are also used to identify in vivo SUMO substrates in yeast and in mammals. Because of low steady-state levels of sumoylation and biases towards abundant targets, identifying sumoylated proteins in vivo can be challenging. The in vitro expression cloning (IVEC) method for SUMO target identification circumvents these challenges and complements the affinity-based approaches. IVEC allows for immediate validation and analysis of substrates through in vitro reconstitution. Furthermore, this method can be easily adapted to identify substrates of specific SUMO ligases.

AB - Covalent modification of proteins by small ubiquitin-like modifier (SUMO) regulates diverse cellular processes. While many SUMO substrates are identified through individual efforts, affinity-based approaches followed by mass spectrometry are also used to identify in vivo SUMO substrates in yeast and in mammals. Because of low steady-state levels of sumoylation and biases towards abundant targets, identifying sumoylated proteins in vivo can be challenging. The in vitro expression cloning (IVEC) method for SUMO target identification circumvents these challenges and complements the affinity-based approaches. IVEC allows for immediate validation and analysis of substrates through in vitro reconstitution. Furthermore, this method can be easily adapted to identify substrates of specific SUMO ligases.

KW - In vitro expression cloning

KW - SUMO

KW - SUMO ligase

KW - Sumoylation

KW - Ubc9

UR - http://www.scopus.com/inward/record.url?scp=59249093852&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=59249093852&partnerID=8YFLogxK

U2 - 10.1007/978-1-59745-566-4_4

DO - 10.1007/978-1-59745-566-4_4

M3 - Chapter

C2 - 19107410

AN - SCOPUS:59249093852

SN - 9781934115800

VL - 497

T3 - Methods in Molecular Biology

SP - 51

EP - 61

BT - Methods in Molecular Biology

ER -