The human selenoprotein VCP-interacting membrane protein (VIMP) is non-globular and harbors a reductase function in an intrinsically disordered region

Lea Cecilie Christensen; Njal Winther Jensen; Andrea Lages Lino Vala; Jurate Kamarauskaite; Linda Christina Johansson; Jakob R. Winther; Kay Hofmann; Kaare Teilum; Lars Ellgaard

doi:10.1074/jbc.M112.346775

The human selenoprotein VCP-interacting membrane protein (VIMP) is non-globular and harbors a reductase function in an intrinsically disordered region

Lea Cecilie Christensen, Njal Winther Jensen, Andrea Lages Lino Vala, Jurate Kamarauskaite, Linda Christina Johansson, Jakob R. Winther, Kay Hofmann, Kaare Teilum, Lars Ellgaard

Biomolecular Sciences

33 Citations (Scopus)

Abstract

The human selenoprotein VIMP (VCP-interacting membrane protein)/SelS (selenoprotein S) localizes to the endoplasmic reticulum (ER) membrane and is involved in the process of ER-associated degradation (ERAD). To date, little is known about the presumed redox activity of VIMP, its structure and how these features might relate to the function of the protein in ERAD. Here, we use the recombinantly expressed cytosolic region of VIMP where the selenocysteine (Sec) in position 188 is replaced with a cysteine (a construct named cVIMP-Cys) to characterize redox and structural properties of the protein. We show that Cys-188 in cVIMP-Cys forms a disulfide bond with Cys-174, consistent with the presence of a Cys174-Sec188 selenosulfide bond in the native sequence. For the disulfide bond in cVIMP-Cys we determined the reduction potential to -200 mV, and showed it to be a good substrate of thioredoxin. Based on a biochemical and structural characterization of cVIMP-Cys using analytical gel filtration, CD and NMR spectroscopy in conjunction with bioinformatics, we propose a comprehensive overall structural model for the cytosolic region of VIMP. The data clearly indicate the N-terminal half to be comprised of two extended α-helices followed by a C-terminal region that is intrinsically disordered. Redox-dependent conformational changes in cVIMP-Cys were observed only in the vicinity of the two Cys residues. Overall, the redox properties observed for cVIMP-Cys are compatible with a function as a reductase, and we speculate that the plasticity of the intrinsically disordered C-terminal region allows the protein to access many different and structurally diverse substrates.

Original language	English
Journal	Journal of Biological Chemistry
Volume	287
Issue number	31
Pages (from-to)	26388-26399
Number of pages	12
ISSN	0021-9258
DOIs	https://doi.org/10.1074/jbc.M112.346775
Publication status	Published - 27 Jul 2012

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10.1074/jbc.M112.346775

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Christensen, L. C., Jensen, N. W., Lages Lino Vala, A., Kamarauskaite, J., Johansson, L. C., Winther, J. R., Hofmann, K., Teilum, K., & Ellgaard, L. (2012). The human selenoprotein VCP-interacting membrane protein (VIMP) is non-globular and harbors a reductase function in an intrinsically disordered region. Journal of Biological Chemistry, 287(31), 26388-26399. https://doi.org/10.1074/jbc.M112.346775

The human selenoprotein VCP-interacting membrane protein (VIMP) is non-globular and harbors a reductase function in an intrinsically disordered region. / Christensen, Lea Cecilie; Jensen, Njal Winther; Lages Lino Vala, Andrea et al.
In: Journal of Biological Chemistry, Vol. 287, No. 31, 27.07.2012, p. 26388-26399.

Research output: Contribution to journal › Journal article › Research › peer-review

Christensen, LC, Jensen, NW, Lages Lino Vala, A, Kamarauskaite, J, Johansson, LC, Winther, JR, Hofmann, K, Teilum, K & Ellgaard, L 2012, 'The human selenoprotein VCP-interacting membrane protein (VIMP) is non-globular and harbors a reductase function in an intrinsically disordered region', Journal of Biological Chemistry, vol. 287, no. 31, pp. 26388-26399. https://doi.org/10.1074/jbc.M112.346775

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title = "The human selenoprotein VCP-interacting membrane protein (VIMP) is non-globular and harbors a reductase function in an intrinsically disordered region",

abstract = "The human selenoprotein VIMP (VCP-interacting membrane protein)/SelS (selenoprotein S) localizes to the endoplasmic reticulum (ER) membrane and is involved in the process of ER-associated degradation (ERAD). To date, little is known about the presumed redox activity of VIMP, its structure and how these features might relate to the function of the protein in ERAD. Here, we use the recombinantly expressed cytosolic region of VIMP where the selenocysteine (Sec) in position 188 is replaced with a cysteine (a construct named cVIMP-Cys) to characterize redox and structural properties of the protein. We show that Cys-188 in cVIMP-Cys forms a disulfide bond with Cys-174, consistent with the presence of a Cys174-Sec188 selenosulfide bond in the native sequence. For the disulfide bond in cVIMP-Cys we determined the reduction potential to -200 mV, and showed it to be a good substrate of thioredoxin. Based on a biochemical and structural characterization of cVIMP-Cys using analytical gel filtration, CD and NMR spectroscopy in conjunction with bioinformatics, we propose a comprehensive overall structural model for the cytosolic region of VIMP. The data clearly indicate the N-terminal half to be comprised of two extended α-helices followed by a C-terminal region that is intrinsically disordered. Redox-dependent conformational changes in cVIMP-Cys were observed only in the vicinity of the two Cys residues. Overall, the redox properties observed for cVIMP-Cys are compatible with a function as a reductase, and we speculate that the plasticity of the intrinsically disordered C-terminal region allows the protein to access many different and structurally diverse substrates.",

author = "Christensen, {Lea Cecilie} and Jensen, {Njal Winther} and {Lages Lino Vala}, Andrea and Jurate Kamarauskaite and Johansson, {Linda Christina} and Winther, {Jakob R.} and Kay Hofmann and Kaare Teilum and Lars Ellgaard",

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T1 - The human selenoprotein VCP-interacting membrane protein (VIMP) is non-globular and harbors a reductase function in an intrinsically disordered region

AU - Christensen, Lea Cecilie

AU - Jensen, Njal Winther

AU - Lages Lino Vala, Andrea

AU - Kamarauskaite, Jurate

AU - Johansson, Linda Christina

AU - Winther, Jakob R.

AU - Hofmann, Kay

AU - Teilum, Kaare

AU - Ellgaard, Lars

PY - 2012/7/27

Y1 - 2012/7/27

N2 - The human selenoprotein VIMP (VCP-interacting membrane protein)/SelS (selenoprotein S) localizes to the endoplasmic reticulum (ER) membrane and is involved in the process of ER-associated degradation (ERAD). To date, little is known about the presumed redox activity of VIMP, its structure and how these features might relate to the function of the protein in ERAD. Here, we use the recombinantly expressed cytosolic region of VIMP where the selenocysteine (Sec) in position 188 is replaced with a cysteine (a construct named cVIMP-Cys) to characterize redox and structural properties of the protein. We show that Cys-188 in cVIMP-Cys forms a disulfide bond with Cys-174, consistent with the presence of a Cys174-Sec188 selenosulfide bond in the native sequence. For the disulfide bond in cVIMP-Cys we determined the reduction potential to -200 mV, and showed it to be a good substrate of thioredoxin. Based on a biochemical and structural characterization of cVIMP-Cys using analytical gel filtration, CD and NMR spectroscopy in conjunction with bioinformatics, we propose a comprehensive overall structural model for the cytosolic region of VIMP. The data clearly indicate the N-terminal half to be comprised of two extended α-helices followed by a C-terminal region that is intrinsically disordered. Redox-dependent conformational changes in cVIMP-Cys were observed only in the vicinity of the two Cys residues. Overall, the redox properties observed for cVIMP-Cys are compatible with a function as a reductase, and we speculate that the plasticity of the intrinsically disordered C-terminal region allows the protein to access many different and structurally diverse substrates.

AB - The human selenoprotein VIMP (VCP-interacting membrane protein)/SelS (selenoprotein S) localizes to the endoplasmic reticulum (ER) membrane and is involved in the process of ER-associated degradation (ERAD). To date, little is known about the presumed redox activity of VIMP, its structure and how these features might relate to the function of the protein in ERAD. Here, we use the recombinantly expressed cytosolic region of VIMP where the selenocysteine (Sec) in position 188 is replaced with a cysteine (a construct named cVIMP-Cys) to characterize redox and structural properties of the protein. We show that Cys-188 in cVIMP-Cys forms a disulfide bond with Cys-174, consistent with the presence of a Cys174-Sec188 selenosulfide bond in the native sequence. For the disulfide bond in cVIMP-Cys we determined the reduction potential to -200 mV, and showed it to be a good substrate of thioredoxin. Based on a biochemical and structural characterization of cVIMP-Cys using analytical gel filtration, CD and NMR spectroscopy in conjunction with bioinformatics, we propose a comprehensive overall structural model for the cytosolic region of VIMP. The data clearly indicate the N-terminal half to be comprised of two extended α-helices followed by a C-terminal region that is intrinsically disordered. Redox-dependent conformational changes in cVIMP-Cys were observed only in the vicinity of the two Cys residues. Overall, the redox properties observed for cVIMP-Cys are compatible with a function as a reductase, and we speculate that the plasticity of the intrinsically disordered C-terminal region allows the protein to access many different and structurally diverse substrates.

U2 - 10.1074/jbc.M112.346775

DO - 10.1074/jbc.M112.346775

M3 - Journal article

C2 - 22700979

SN - 0021-9258

VL - 287

SP - 26388

EP - 26399

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 31

ER -