Expression, purification and insights into structure and folding of the ADAM22 pro domain.

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Expression, purification and insights into structure and folding of the ADAM22 pro domain. / Sørensen, Hans Peter; Jacobsen, Jonas; Nielbo, Steen; Poulsen, Flemming M; Wewer, Ulla M.

In: Protein Expression and Purification, Vol. 61, No. 2, 2008, p. 175-83.

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

Harvard

Sørensen, HP, Jacobsen, J, Nielbo, S, Poulsen, FM & Wewer, UM 2008, 'Expression, purification and insights into structure and folding of the ADAM22 pro domain.', Protein Expression and Purification, vol. 61, no. 2, pp. 175-83. https://doi.org/10.1016/j.pep.2008.05.020

APA

Sørensen, H. P., Jacobsen, J., Nielbo, S., Poulsen, F. M., & Wewer, U. M. (2008). Expression, purification and insights into structure and folding of the ADAM22 pro domain. Protein Expression and Purification, 61(2), 175-83. https://doi.org/10.1016/j.pep.2008.05.020

Vancouver

Sørensen HP, Jacobsen J, Nielbo S, Poulsen FM, Wewer UM. Expression, purification and insights into structure and folding of the ADAM22 pro domain. Protein Expression and Purification. 2008;61(2):175-83. https://doi.org/10.1016/j.pep.2008.05.020

Author

Sørensen, Hans Peter ; Jacobsen, Jonas ; Nielbo, Steen ; Poulsen, Flemming M ; Wewer, Ulla M. / Expression, purification and insights into structure and folding of the ADAM22 pro domain. In: Protein Expression and Purification. 2008 ; Vol. 61, No. 2. pp. 175-83.

Bibtex

@article{61a875e0ac0f11ddb5e9000ea68e967b,
title = "Expression, purification and insights into structure and folding of the ADAM22 pro domain.",
abstract = "The ADAMs (a disintegrin and metalloproteases) are an important class of enzymes in the regulation of human disease. The pro domains of ADAMs are responsible for the latency and secretion of mature enzymes. Unlike other metzincins, ADAM pro domains remain bound to the mature enzyme after secretion. To understand the functions of human ADAM pro domains and to determine three-dimensional structures, we have screened promising targets for expression and purification properties when using Escherichia coli as the host. The pro domain of ADAM22 (ADAM22-P) expressed in E. coli was folded, as determined by CD and NMR spectroscopy. An ADAM22-P fragment encoding residues 26-199 could be expressed in high amounts, remained soluble above 1 mM, and was suitable for structural studies by NMR spectroscopy. CD spectroscopy and predictions suggest that the secondary structure in ADAM22-P consists of beta-strands. Furthermore, our data indicate that the pro domains of ADAMs are expressed as two subdomains. The most N-terminal subdomain (ADAM22-P(N)) was found to be susceptible to proteolysis and was required for folding stability of the second subdomain (ADAM22-P(C)).",
author = "S{\o}rensen, {Hans Peter} and Jonas Jacobsen and Steen Nielbo and Poulsen, {Flemming M} and Wewer, {Ulla M}",
year = "2008",
doi = "10.1016/j.pep.2008.05.020",
language = "English",
volume = "61",
pages = "175--83",
journal = "Protein Expression and Purification",
issn = "1046-5928",
publisher = "Academic Press",
number = "2",

}

RIS

TY - JOUR

T1 - Expression, purification and insights into structure and folding of the ADAM22 pro domain.

AU - Sørensen, Hans Peter

AU - Jacobsen, Jonas

AU - Nielbo, Steen

AU - Poulsen, Flemming M

AU - Wewer, Ulla M

PY - 2008

Y1 - 2008

N2 - The ADAMs (a disintegrin and metalloproteases) are an important class of enzymes in the regulation of human disease. The pro domains of ADAMs are responsible for the latency and secretion of mature enzymes. Unlike other metzincins, ADAM pro domains remain bound to the mature enzyme after secretion. To understand the functions of human ADAM pro domains and to determine three-dimensional structures, we have screened promising targets for expression and purification properties when using Escherichia coli as the host. The pro domain of ADAM22 (ADAM22-P) expressed in E. coli was folded, as determined by CD and NMR spectroscopy. An ADAM22-P fragment encoding residues 26-199 could be expressed in high amounts, remained soluble above 1 mM, and was suitable for structural studies by NMR spectroscopy. CD spectroscopy and predictions suggest that the secondary structure in ADAM22-P consists of beta-strands. Furthermore, our data indicate that the pro domains of ADAMs are expressed as two subdomains. The most N-terminal subdomain (ADAM22-P(N)) was found to be susceptible to proteolysis and was required for folding stability of the second subdomain (ADAM22-P(C)).

AB - The ADAMs (a disintegrin and metalloproteases) are an important class of enzymes in the regulation of human disease. The pro domains of ADAMs are responsible for the latency and secretion of mature enzymes. Unlike other metzincins, ADAM pro domains remain bound to the mature enzyme after secretion. To understand the functions of human ADAM pro domains and to determine three-dimensional structures, we have screened promising targets for expression and purification properties when using Escherichia coli as the host. The pro domain of ADAM22 (ADAM22-P) expressed in E. coli was folded, as determined by CD and NMR spectroscopy. An ADAM22-P fragment encoding residues 26-199 could be expressed in high amounts, remained soluble above 1 mM, and was suitable for structural studies by NMR spectroscopy. CD spectroscopy and predictions suggest that the secondary structure in ADAM22-P consists of beta-strands. Furthermore, our data indicate that the pro domains of ADAMs are expressed as two subdomains. The most N-terminal subdomain (ADAM22-P(N)) was found to be susceptible to proteolysis and was required for folding stability of the second subdomain (ADAM22-P(C)).

U2 - 10.1016/j.pep.2008.05.020

DO - 10.1016/j.pep.2008.05.020

M3 - Journal article

C2 - 18593599

VL - 61

SP - 175

EP - 183

JO - Protein Expression and Purification

JF - Protein Expression and Purification

SN - 1046-5928

IS - 2

ER -

ID: 8443587