title: Covalent Capturing of Transient SUMO-SIM Interactions Using Unnatural Amino Acid Mutagenesis and Photocrosslinking.
abstract: The weak interaction between the post-translational modifier SUMO (small ubiquitin-like modifier) and proteins containing the SUMO-interacting motif (SIM) poses limitations to the identification of interaction partners of SUMOylated proteins and to the mapping of the interfaces. To overcome these limitations, genetically encoded photocrosslinker amino acids were incorporated close to the SIM-interaction groove in human SUMO1. UV irradiation resulted in the desired covalent crosslinks both in a purified protein environment and in cell extracts.
authors.affiliation: Department of Chemistry and Pharmacy, Institute of Biochemistry, University of Muenster, Wilhelm-Klemm-Str. 2, 48149, Münster, Germany.
authors.forename: Kim F
authors.initials: KF
authors.lastname: Taupitz
authors.affiliation: Department of Chemistry and Pharmacy, Institute of Biochemistry, University of Muenster, Wilhelm-Klemm-Str. 2, 48149, Münster, Germany.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Dörner
authors.affiliation: Department of Chemistry and Pharmacy, Institute of Biochemistry, University of Muenster, Wilhelm-Klemm-Str. 2, 48149, Münster, Germany.
authors.forename: Henning D
authors.identifier: http://orcid.org/0000-0002-3385-0234
authors.initials: HD
authors.lastname: Mootz
mesh_terms: D020816:Amino Acid Motifs; D000596:Amino Acids / Q000378:metabolism; D051792:Basic Helix-Loop-Helix Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D016296:Mutagenesis; D010455:Peptides / Q000032:analysis; D011485:Protein Binding; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D025842:SUMO-1 Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D014466:Ultraviolet Rays
chemical_list: D000596:Amino Acids; D051792:Basic Helix-Loop-Helix Proteins; D010455:Peptides; D012097:Repressor Proteins; D025842:SUMO-1 Protein; C491094:SIM1 protein, human
keywords: SUMO; crosslinker; genetic code expansion; post-translational modification; protein interaction