Recombinant human CIS2 (SOCS2) protein: Subcloning, expression, purification, and characterization

Eva Biener; Sarah Maurice; Yael Sandowski; Yael Cohen; Eugene E. Gusakowsky; Robert Hooghe; Akihiko Yoshimura; Oded Livnah; Arieh Gertler

doi:10.1016/S1046-5928(02)00013-X

Recombinant human CIS2 (SOCS2) protein: Subcloning, expression, purification, and characterization

Eva Biener, Sarah Maurice, Yael Sandowski, Yael Cohen, Eugene E. Gusakowsky, Robert Hooghe, Akihiko Yoshimura, Oded Livnah, Arieh Gertler^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

The 1x myc-tagged cDNA encoding for human CIS2 protein was subcloned into a pET-29a+ vector in order to express and produce a recombinant S-peptide tagged and 1x myc-tagged protein in Escherichia coli BL21(DE3). The constitutively expressed protein was isolated from inclusion bodies by a simple solubilization-renaturation procedure and purified by anion-exchange chromatography on Q-Sepharose. The recombinant form was found to be pure and monomeric as judged by both SDS-PAGE and gel-filtration chromatography and its biological activity was proven by its ability to bind to the tyrosine-phosphorylated cytosolic fragment of human growth hormone receptor fused to glutathione-S-transferase. Recombinant CIS2 was compared by biochemical, immunological, and molecular methods to the CIS2 protein expressed in eukaryotic cells. This report describes the first substantial production of biologically active recombinant human CIS2.

Original language	English
Pages (from-to)	305-312
Number of pages	8
Journal	Protein Expression and Purification
Volume	25
Issue number	2
DOIs	https://doi.org/10.1016/S1046-5928(02)00013-X
State	Published - 2002

Bibliographical note

Funding Information:
We thank Dr. Nils Billestrup from Hagedorn Research Institute (Denmark) for the gift of bacterial plasmid encoding the GST–GHR. This work was supported by the Israeli Science Foundation (Grant 460/99) to A. Gertler and O. Livnah and partially supported by a grant (to B. Velkeniers) from the Ministry of Scientific Research of the Brussels-Capital Region.

Keywords

CIS2
Growth hormone receptor
Recombinant proteins

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10.1016/S1046-5928(02)00013-X

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title = "Recombinant human CIS2 (SOCS2) protein: Subcloning, expression, purification, and characterization",

abstract = "The 1x myc-tagged cDNA encoding for human CIS2 protein was subcloned into a pET-29a+ vector in order to express and produce a recombinant S-peptide tagged and 1x myc-tagged protein in Escherichia coli BL21(DE3). The constitutively expressed protein was isolated from inclusion bodies by a simple solubilization-renaturation procedure and purified by anion-exchange chromatography on Q-Sepharose. The recombinant form was found to be pure and monomeric as judged by both SDS-PAGE and gel-filtration chromatography and its biological activity was proven by its ability to bind to the tyrosine-phosphorylated cytosolic fragment of human growth hormone receptor fused to glutathione-S-transferase. Recombinant CIS2 was compared by biochemical, immunological, and molecular methods to the CIS2 protein expressed in eukaryotic cells. This report describes the first substantial production of biologically active recombinant human CIS2.",

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author = "Eva Biener and Sarah Maurice and Yael Sandowski and Yael Cohen and Gusakowsky, {Eugene E.} and Robert Hooghe and Akihiko Yoshimura and Oded Livnah and Arieh Gertler",

note = "Funding Information: We thank Dr. Nils Billestrup from Hagedorn Research Institute (Denmark) for the gift of bacterial plasmid encoding the GST–GHR. This work was supported by the Israeli Science Foundation (Grant 460/99) to A. Gertler and O. Livnah and partially supported by a grant (to B. Velkeniers) from the Ministry of Scientific Research of the Brussels-Capital Region.",

year = "2002",

doi = "10.1016/S1046-5928(02)00013-X",

language = "אנגלית",

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T2 - Subcloning, expression, purification, and characterization

AU - Biener, Eva

AU - Maurice, Sarah

AU - Sandowski, Yael

AU - Cohen, Yael

AU - Gusakowsky, Eugene E.

AU - Hooghe, Robert

AU - Yoshimura, Akihiko

AU - Livnah, Oded

AU - Gertler, Arieh

N1 - Funding Information: We thank Dr. Nils Billestrup from Hagedorn Research Institute (Denmark) for the gift of bacterial plasmid encoding the GST–GHR. This work was supported by the Israeli Science Foundation (Grant 460/99) to A. Gertler and O. Livnah and partially supported by a grant (to B. Velkeniers) from the Ministry of Scientific Research of the Brussels-Capital Region.

PY - 2002

Y1 - 2002

N2 - The 1x myc-tagged cDNA encoding for human CIS2 protein was subcloned into a pET-29a+ vector in order to express and produce a recombinant S-peptide tagged and 1x myc-tagged protein in Escherichia coli BL21(DE3). The constitutively expressed protein was isolated from inclusion bodies by a simple solubilization-renaturation procedure and purified by anion-exchange chromatography on Q-Sepharose. The recombinant form was found to be pure and monomeric as judged by both SDS-PAGE and gel-filtration chromatography and its biological activity was proven by its ability to bind to the tyrosine-phosphorylated cytosolic fragment of human growth hormone receptor fused to glutathione-S-transferase. Recombinant CIS2 was compared by biochemical, immunological, and molecular methods to the CIS2 protein expressed in eukaryotic cells. This report describes the first substantial production of biologically active recombinant human CIS2.

AB - The 1x myc-tagged cDNA encoding for human CIS2 protein was subcloned into a pET-29a+ vector in order to express and produce a recombinant S-peptide tagged and 1x myc-tagged protein in Escherichia coli BL21(DE3). The constitutively expressed protein was isolated from inclusion bodies by a simple solubilization-renaturation procedure and purified by anion-exchange chromatography on Q-Sepharose. The recombinant form was found to be pure and monomeric as judged by both SDS-PAGE and gel-filtration chromatography and its biological activity was proven by its ability to bind to the tyrosine-phosphorylated cytosolic fragment of human growth hormone receptor fused to glutathione-S-transferase. Recombinant CIS2 was compared by biochemical, immunological, and molecular methods to the CIS2 protein expressed in eukaryotic cells. This report describes the first substantial production of biologically active recombinant human CIS2.

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Recombinant human CIS2 (SOCS2) protein: Subcloning, expression, purification, and characterization

Abstract

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Keywords

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