Swapping Small Ubiquitin-like Modifier (SUMO) Isoform Specificity of SUMO Proteases SENP6 and SENP7

Alegre, Kamela O.; Reverter, David

doi:10.1074/jbc.m111.268847

Cited by 35 publications

(39 citation statements)

References 35 publications

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“…26,27 Both enzymes, however, have demonstrated activity in deconjugating reactions with SUMO substrates, in particular against polySUMOylated chains, and show isoform specificity for SUMO2/3 over SUMO1. [26][27][28] As mentioned before, SENP6 and SENP7 are characterized by the presence of long insertions in their catalytic domains. In vitro deconjugation assays with constructs lacking these inserts, however, show similar functional activities, leaving the relevance of these large insertions to be elucidated.…”

Section: Introductionmentioning

confidence: 99%

“…26,27 Of special interest was the relevance of the Loop1 insertion for activity, unique to SENP6 and SENP7 proteases, which presents only a single amino acid substitution between these two proteases (alanine and threonine for SENP6 and SENP7, respectively) ( Fig. 1).…”

Section: Production Of the Senp2-loop1 Chimeramentioning

confidence: 99%

“…The crystal structure of the apo SENP7 revealed that Loop1 is structured and its deletion produced marked defects in the proteolytic activities of both SENP6 and SENP7. 26,27 To get direct structural details of the Loop1 interface with SUMO2/3, we introduced Loop1 into the catalytic domain of SENP2. The SENP6 Loop1 sequence was inserted at position 392 of SENP2 Figure 1.…”

Section: Production Of the Senp2-loop1 Chimeramentioning

confidence: 99%

“…24,25 SENP6 and SENP7 show little in vitro activity in processing reactions of SUMO precursors. 26,27 Both enzymes, however, have demonstrated activity in deconjugating reactions with SUMO substrates, in particular against polySUMOylated chains, and show isoform specificity for SUMO2/3 over SUMO1. [26][27][28] As mentioned before, SENP6 and SENP7 are characterized by the presence of long insertions in their catalytic domains.…”

Section: Introductionmentioning

confidence: 99%

“…In vitro deconjugation assays with constructs lacking these inserts, however, show similar functional activities, leaving the relevance of these large insertions to be elucidated. 26,27 Nevertheless, Loop1 insertion, a short 8-residue insertion unique to SENP6 and SENP7, is essential for the in vitro proteolytic activity and is determinant for the SUMO2/3 isoform specificity of SENP6 and SENP7.…”

Section: Introductionmentioning

confidence: 99%

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Structural insights into the SENP6 Loop1 structure in complex with SUMO2

Alegre

Reverter

2014

Protein Science

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The SENP proteases regulate the SUMO conjugates in the cell by cleaving SUMO from target proteins. SENP6 and SENP7 are the most divergent members of the SENP/ULP protease family in humans by the presence of insertions in their catalytic domains. Loop1 insertion is determinant for the SUMO2/3 activity and specificity on SENP6 and SENP7. To gain structural insights into the role of Loop1, we have designed a chimeric SENP2 with the insertion of Loop1 into its sequence. The structure of SENP2-Loop1 in complex with SUMO2 was solved at 2.15 Å resolution, and reveals the details of an interface exclusive to SENP6/7 and the formation of unique contacts between both proteins. Interestingly, functional data with SUMO substrates showed an increase of the proteolytic activity in the SENP2-Loop1 chimera for diSUMO2 and polySUMO2 substrates.

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Section: Introductionmentioning

confidence: 99%

Section: Production Of the Senp2-loop1 Chimeramentioning

confidence: 99%

Section: Production Of the Senp2-loop1 Chimeramentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Structural insights into the SENP6 Loop1 structure in complex with SUMO2

Alegre

Reverter

2014

Protein Science

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In‐Plate Chemical Synthesis of Isopeptide‐Linked SUMOylated Peptide Fluorescence Polarization Reagents for High‐Throughput Screening of SENP Preferences

et al. 2022

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Small ubiquitin-like modifiers (SUMOs) are conjugated to protein substrates in cells to regulate their function. The attachment of SUMO family members SUMO1-3 to substrate proteins is reversed by specific isopeptidases called SENPs (sentrin-specific protease). Whereas SENPs are SUMO-isoform or linkage type specific, comprehensive analysis is missing. Furthermore, the underlying mechanism of SENP linkage specificity remains unclear. We present a high-throughput synthesis of 83 isopeptide-linked SUMO-based fluorescence polarization reagents to study enzyme preferences. The assay reagents were synthesized via a native chemical ligation-desulfurization protocol between 11-mer peptides containing a γ-thiolysine and a SUMO3 thioester. Subsequently, five recombinantly expressed SENPs were screened using these assay reagents to reveal their deconjugation activity and substrate preferences. In general, we observed that SENP1 is the most active and nonselective SENP while SENP6 and SENP7 show the least activity. Furthermore, SENPs differentially process peptides derived from SUMO1-3, who form a minimalistic representation of diSUMO chains. To validate our findings, five distinct isopeptide-linked diSUMO chains were chemically synthesized and proteolysis was monitored using a gel-based read-out.

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Wrestling with stress: Roles of protein SUMOylation and deSUMOylation in cell stress response

2014

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How cell fate is determined following extreme stress is a core question in cell biology. This is particularly important in the brain where neuronal death following ischemic stroke is a major cause of disability. Over the last few years it has emerged that the SUMOylation status of an increasing number of substrate proteins plays a crucial role in cellular responses to environmental and metabolic stress. SUMOylation is a post-translational modification in which the 97-residue protein, SUMO (Small Ubiquitinrelated MOdifier) is covalently attached to specific lysine residues in a target protein. Despite being covalent, it is a highly transient modification because of the actions of deSUMOylation enzymes, so SUMO conjugation acts as a rapidly reversible switch that can promote or inhibit protein interactions with the substrate protein.Overall, it appears that increased SUMOylation represents a cellular protective response. Here we discuss recent progress toward understanding the mechanisms, pathways, and roles of SUMOylation during and after severe metabolic stress. V C 2014 IUBMB Life, 66(2): [71][72][73][74][75][76][77] 2014

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Swapping Small Ubiquitin-like Modifier (SUMO) Isoform Specificity of SUMO Proteases SENP6 and SENP7

Cited by 35 publications

References 35 publications

Structural insights into the SENP6 Loop1 structure in complex with SUMO2

Structural insights into the SENP6 Loop1 structure in complex with SUMO2

In‐Plate Chemical Synthesis of Isopeptide‐Linked SUMOylated Peptide Fluorescence Polarization Reagents for High‐Throughput Screening of SENP Preferences

Wrestling with stress: Roles of protein SUMOylation and deSUMOylation in cell stress response

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