Purification and biochemical characterization of affinity-tagged Musca domestica acetylcholinesterase secreted from baculovirus infected cells

Graham D. Moores; Martin S. Williamson; Medard Hadonou; Alan L. Devonshire; Kari Kataja; Juha Uitto; Christer Oker-Blom

doi:10.1007/978-1-4899-1540-5_145

Purification and biochemical characterization of affinity-tagged Musca domestica acetylcholinesterase secreted from baculovirus infected cells

Graham D. Moores, Martin S. Williamson, Medard Hadonou, Alan L. Devonshire, Kari Kataja, Juha Uitto, Christer Oker-Blom

VTT Technical Research Centre of Finland

Research output: Chapter in Book/Report/Conference proceeding › Conference article in proceedings › Scientific › peer-review

Abstract

The baculovirus expression system was employed for production of a modified, FLAG-epitope tagged Musca domestica acetylcholinestearse (AChE) in Sf9 cells. The signal peptide of ecdysteroid UDP-glycosyltransferase (EGT) of Autographa californica nuclear polyhedrosis virus was utilized for translocation of the protein into the secretory pathway of the insect cell. Further, secretion was enabled by truncation of its hydrophobic domain. The protein was produced in large scale, and purified from the extra cellular space of the infected cell cultures by immunoaffinity chromatography using the mouse monoclonal Ml antibody. Up to 30 mg of biologically active protein was produced per liter of cell culture.

Original language	English
Title of host publication	Structure and Function of Cholinesterases and Related Proteins
Editors	Bhupendra P. Doctor, Palmer Taylor, Daniel M. Quinn, Richard L. Rotundo, Mary K. Gentry
Place of Publication	New York
Publisher	Plenum
Pages	547
ISBN (Electronic)	978-1-4899-1540-5
ISBN (Print)	978-1-4899-1542-9, 978-0-306-46050-0
DOIs	https://doi.org/10.1007/978-1-4899-1540-5_145
Publication status	Published - 1998
MoE publication type	A4 Article in a conference publication
Event	6th International Meeting on Cholinesterases and Related Proteins: Cholinesterases '98 - San Diego, United States Duration: 1 Mar 1998 → …

Conference

Conference	6th International Meeting on Cholinesterases and Related Proteins
Country/Territory	United States
City	San Diego
Period	1/03/98 → …

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10.1007/978-1-4899-1540-5_145

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Moores, G. D., Williamson, M. S., Hadonou, M., Devonshire, A. L., Kataja, K., Uitto, J., & Oker-Blom, C. (1998). Purification and biochemical characterization of affinity-tagged Musca domestica acetylcholinesterase secreted from baculovirus infected cells. In B. P. Doctor, P. Taylor, D. M. Quinn, R. L. Rotundo, & M. K. Gentry (Eds.), Structure and Function of Cholinesterases and Related Proteins (pp. 547). Plenum. https://doi.org/10.1007/978-1-4899-1540-5_145

Moores, Graham D. ; Williamson, Martin S. ; Hadonou, Medard et al. / Purification and biochemical characterization of affinity-tagged Musca domestica acetylcholinesterase secreted from baculovirus infected cells. Structure and Function of Cholinesterases and Related Proteins. editor / Bhupendra P. Doctor ; Palmer Taylor ; Daniel M. Quinn ; Richard L. Rotundo ; Mary K. Gentry. New York : Plenum, 1998. pp. 547

@inproceedings{cd72921f16124126ac1901bac8570595,

title = "Purification and biochemical characterization of affinity-tagged Musca domestica acetylcholinesterase secreted from baculovirus infected cells",

abstract = "The baculovirus expression system was employed for production of a modified, FLAG-epitope tagged Musca domestica acetylcholinestearse (AChE) in Sf9 cells. The signal peptide of ecdysteroid UDP-glycosyltransferase (EGT) of Autographa californica nuclear polyhedrosis virus was utilized for translocation of the protein into the secretory pathway of the insect cell. Further, secretion was enabled by truncation of its hydrophobic domain. The protein was produced in large scale, and purified from the extra cellular space of the infected cell cultures by immunoaffinity chromatography using the mouse monoclonal Ml antibody. Up to 30 mg of biologically active protein was produced per liter of cell culture.",

author = "Moores, \{Graham D.\} and Williamson, \{Martin S.\} and Medard Hadonou and Devonshire, \{Alan L.\} and Kari Kataja and Juha Uitto and Christer Oker-Blom",

year = "1998",

doi = "10.1007/978-1-4899-1540-5\_145",

language = "English",

isbn = "978-1-4899-1542-9",

pages = "547",

editor = "Doctor, \{Bhupendra P.\} and Palmer Taylor and Quinn, \{Daniel M.\} and Rotundo, \{Richard L.\} and Gentry, \{Mary K.\}",

booktitle = "Structure and Function of Cholinesterases and Related Proteins",

publisher = "Plenum",

address = "United States",

note = "6th International Meeting on Cholinesterases and Related Proteins : Cholinesterases '98 ; Conference date: 01-03-1998",

}

Moores, GD, Williamson, MS, Hadonou, M, Devonshire, AL, Kataja, K, Uitto, J & Oker-Blom, C 1998, Purification and biochemical characterization of affinity-tagged Musca domestica acetylcholinesterase secreted from baculovirus infected cells. in BP Doctor, P Taylor, DM Quinn, RL Rotundo & MK Gentry (eds), Structure and Function of Cholinesterases and Related Proteins. Plenum, New York, pp. 547, 6th International Meeting on Cholinesterases and Related Proteins, San Diego, California, United States, 1/03/98. https://doi.org/10.1007/978-1-4899-1540-5_145

Purification and biochemical characterization of affinity-tagged Musca domestica acetylcholinesterase secreted from baculovirus infected cells. / Moores, Graham D.; Williamson, Martin S.; Hadonou, Medard et al.
Structure and Function of Cholinesterases and Related Proteins. ed. / Bhupendra P. Doctor; Palmer Taylor; Daniel M. Quinn; Richard L. Rotundo; Mary K. Gentry. New York: Plenum, 1998. p. 547.

Research output: Chapter in Book/Report/Conference proceeding › Conference article in proceedings › Scientific › peer-review

TY - GEN

T1 - Purification and biochemical characterization of affinity-tagged Musca domestica acetylcholinesterase secreted from baculovirus infected cells

AU - Moores, Graham D.

AU - Williamson, Martin S.

AU - Hadonou, Medard

AU - Devonshire, Alan L.

AU - Kataja, Kari

AU - Uitto, Juha

AU - Oker-Blom, Christer

PY - 1998

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N2 - The baculovirus expression system was employed for production of a modified, FLAG-epitope tagged Musca domestica acetylcholinestearse (AChE) in Sf9 cells. The signal peptide of ecdysteroid UDP-glycosyltransferase (EGT) of Autographa californica nuclear polyhedrosis virus was utilized for translocation of the protein into the secretory pathway of the insect cell. Further, secretion was enabled by truncation of its hydrophobic domain. The protein was produced in large scale, and purified from the extra cellular space of the infected cell cultures by immunoaffinity chromatography using the mouse monoclonal Ml antibody. Up to 30 mg of biologically active protein was produced per liter of cell culture.

AB - The baculovirus expression system was employed for production of a modified, FLAG-epitope tagged Musca domestica acetylcholinestearse (AChE) in Sf9 cells. The signal peptide of ecdysteroid UDP-glycosyltransferase (EGT) of Autographa californica nuclear polyhedrosis virus was utilized for translocation of the protein into the secretory pathway of the insect cell. Further, secretion was enabled by truncation of its hydrophobic domain. The protein was produced in large scale, and purified from the extra cellular space of the infected cell cultures by immunoaffinity chromatography using the mouse monoclonal Ml antibody. Up to 30 mg of biologically active protein was produced per liter of cell culture.

U2 - 10.1007/978-1-4899-1540-5_145

DO - 10.1007/978-1-4899-1540-5_145

M3 - Conference article in proceedings

SN - 978-1-4899-1542-9

SN - 978-0-306-46050-0

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BT - Structure and Function of Cholinesterases and Related Proteins

A2 - Doctor, Bhupendra P.

A2 - Taylor, Palmer

A2 - Quinn, Daniel M.

A2 - Rotundo, Richard L.

A2 - Gentry, Mary K.

PB - Plenum

CY - New York

T2 - 6th International Meeting on Cholinesterases and Related Proteins

Y2 - 1 March 1998

ER -

Moores GD, Williamson MS, Hadonou M, Devonshire AL, Kataja K, Uitto J et al. Purification and biochemical characterization of affinity-tagged Musca domestica acetylcholinesterase secreted from baculovirus infected cells. In Doctor BP, Taylor P, Quinn DM, Rotundo RL, Gentry MK, editors, Structure and Function of Cholinesterases and Related Proteins. New York: Plenum. 1998. p. 547 doi: 10.1007/978-1-4899-1540-5_145

Purification and biochemical characterization of affinity-tagged Musca domestica acetylcholinesterase secreted from baculovirus infected cells

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