SUMO (small ubiquitin-related modifier) is a member of the ubiquitin-like protein family that regulates cellular function of a variety of target proteins. SUMO proteins are expressed as their precursor forms. Cleavage of the residues after the ‘GG’ region of these precursors by SUMO-specific proteases in maturation is a prerequisite for subsequent sumoylation. To understand further this proteolytic processing, we expressed and purified SENP1 (sentrin-specific protease 1), one of the SUMO-specific proteases, using an Escherichia coli expression system. We show that SENP1 is capable of processing all SUMO-1, -2 and -3 in vitro; however, the proteolytic efficiency of SUMO-1 is the highest followed by SUMO-2 and -3. We demonstrate further that the catalytic domain of SENP1 (SENP1C) alone can determine the substrate specificity towards SUMO-1, -2 and -3. Replacement of the C-terminal fragments after the ‘GG’ region of SUMO-1 and -2 precursors with that of the SUMO-3, indicates that the C-terminal fragment is essential for efficient maturation. In mutagenesis analysis, we further map two residues immediately after the ‘GG’ region, which determine the differential maturation. Distinct patterns of tissue distribution of SENP1, SUMO-1, -2 and -3 are characterized. Taken together, we suggest that the observed differential maturation process has its physiological significance in the regulation of the sumoylation pathway.
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Research Article|
February 22 2005
Mapping residues of SUMO precursors essential in differential maturation by SUMO-specific protease, SENP1
Zheng XU;
Zheng XU
1Department of Biochemistry, Faculty of Science, The Chinese University of Hong Kong, Shatin, Hong Kong
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Shannon W. N. AU
Shannon W. N. AU
1
1Department of Biochemistry, Faculty of Science, The Chinese University of Hong Kong, Shatin, Hong Kong
1To whom correspondence should be addressed (email Shannon-au@cuhk.edu.hk).
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Publisher: Portland Press Ltd
Received:
July 19 2004
Revision Received:
October 04 2004
Accepted:
October 15 2004
Accepted Manuscript online:
October 18 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2005
Biochem J (2005) 386 (2): 325–330.
Article history
Received:
July 19 2004
Revision Received:
October 04 2004
Accepted:
October 15 2004
Accepted Manuscript online:
October 18 2004
Citation
Zheng XU, Shannon W. N. AU; Mapping residues of SUMO precursors essential in differential maturation by SUMO-specific protease, SENP1. Biochem J 1 March 2005; 386 (2): 325–330. doi: https://doi.org/10.1042/BJ20041210
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