title: Insights into high affinity small ubiquitin-like modifier (SUMO) recognition by SUMO-interacting motifs (SIMs) revealed by a combination of NMR and peptide array analysis.
abstract: The small ubiquitin-like modifiers (SUMOs) regulate many essential cellular functions. Only one type of SUMO-interacting motif (SIM) has been identified that can extend the β-sheet of SUMO as either a parallel or an antiparallel strand. The molecular determinants of the bound orientation and paralogue specificity of a SIM are unclear. To address this question, we have conducted structural studies of SUMO1 in complex with a SUMO1-specific SIM that binds to SUMO1 with high affinity without post-translational modifications using nuclear magnetic resonance methods. In addition, the SIM sequence requirements have been investigated by peptide arrays in comparison with another high affinity SIM that binds in the opposing orientation. We found that antiparallel binding SIMs tolerate more diverse sequences, whereas the parallel binding SIMs prefer the more strict sequences consisting of (I/V)DLT that have a preference in high affinity SUMO2 and -3 binding. Comparison of two high affinity SUMO1-binding SIMs that bind in opposing orientations has revealed common SUMO1-specific interactions needed for high affinity binding. This study has significantly advanced our understanding of the molecular determinants underlining SUMO-SIM recognition.
authors.affiliation: Department of Molecular Medicine, Beckman Research Institute of City of Hope, Duarte, California 91010, USA.
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Namanja
authors.forename: Yi-Jia
authors.initials: YJ
authors.lastname: Li
authors.forename: Yang
authors.initials: Y
authors.lastname: Su
authors.forename: Steven
authors.initials: S
authors.lastname: Wong
authors.forename: Jingjun
authors.initials: J
authors.lastname: Lu
authors.forename: Loren T
authors.initials: LT
authors.lastname: Colson
authors.forename: Chenggang
authors.initials: C
authors.lastname: Wu
authors.forename: Shawn S C
authors.initials: SS
authors.lastname: Li
authors.forename: Yuan
authors.initials: Y
authors.lastname: Chen
mesh_terms: D020816:Amino Acid Motifs; D045744:Cell Line, Tumor; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular; D025842:SUMO-1 Protein / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D025841:Small Ubiquitin-Related Modifier Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014452:Ubiquitins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D025842:SUMO-1 Protein; C495501:SUMO1 protein, human; C117909:SUMO2 protein, human; C492189:SUMO3 protein, human; D025841:Small Ubiquitin-Related Modifier Proteins; D014452:Ubiquitins