Multiprotein expression strategy for structural biology of eukaryotic complexes

Daniel J Fitzgerald; Christiane Schaffitzel; Philipp Berger; Ralf Wellinger; Christoph Bieniossek; Timothy J Richmond; Imre Berger

doi:10.1016/j.str.2007.01.016

Multiprotein expression strategy for structural biology of eukaryotic complexes

Daniel J Fitzgerald, Christiane Schaffitzel, Philipp Berger, Ralf Wellinger, Christoph Bieniossek, Timothy J Richmond, Imre Berger

School of Biochemistry

Research output: Contribution to journal › Article (Academic Journal) › peer-review

48 Citations (Scopus)

Abstract

The concept of the cell as a collection of multisubunit protein machines is emerging as a cornerstone of modern biology, and molecular-level study of these machines in most cases will require recombinant production. Here, we present and validate a strategy to rapidly produce, permutate, and posttranslationally modify large, eukaryotic multiprotein complexes by using DNA recombination in a process that is fully automatable. Parallel production of 12 protein complex variants within a period of weeks resulted in specimens of sufficient quantity and homogeneity for structural biology applications.

Original language	English
Pages (from-to)	275-9
Number of pages	5
Journal	Structure
Volume	15
Issue number	3
DOIs	https://doi.org/10.1016/j.str.2007.01.016
Publication status	Published - Mar 2007

Keywords

Animals
Baculoviridae
Cell Line
Escherichia coli
Genetic Vectors
Humans
Multiprotein Complexes
Spodoptera

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title = "Multiprotein expression strategy for structural biology of eukaryotic complexes",

abstract = "The concept of the cell as a collection of multisubunit protein machines is emerging as a cornerstone of modern biology, and molecular-level study of these machines in most cases will require recombinant production. Here, we present and validate a strategy to rapidly produce, permutate, and posttranslationally modify large, eukaryotic multiprotein complexes by using DNA recombination in a process that is fully automatable. Parallel production of 12 protein complex variants within a period of weeks resulted in specimens of sufficient quantity and homogeneity for structural biology applications.",

keywords = "Animals, Baculoviridae, Cell Line, Escherichia coli, Genetic Vectors, Humans, Multiprotein Complexes, Spodoptera",

author = "Fitzgerald, \{Daniel J\} and Christiane Schaffitzel and Philipp Berger and Ralf Wellinger and Christoph Bieniossek and Richmond, \{Timothy J\} and Imre Berger",

year = "2007",

month = mar,

doi = "10.1016/j.str.2007.01.016",

language = "English",

volume = "15",

pages = "275--9",

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T1 - Multiprotein expression strategy for structural biology of eukaryotic complexes

AU - Fitzgerald, Daniel J

AU - Schaffitzel, Christiane

AU - Berger, Philipp

AU - Wellinger, Ralf

AU - Bieniossek, Christoph

AU - Richmond, Timothy J

AU - Berger, Imre

PY - 2007/3

Y1 - 2007/3

N2 - The concept of the cell as a collection of multisubunit protein machines is emerging as a cornerstone of modern biology, and molecular-level study of these machines in most cases will require recombinant production. Here, we present and validate a strategy to rapidly produce, permutate, and posttranslationally modify large, eukaryotic multiprotein complexes by using DNA recombination in a process that is fully automatable. Parallel production of 12 protein complex variants within a period of weeks resulted in specimens of sufficient quantity and homogeneity for structural biology applications.

AB - The concept of the cell as a collection of multisubunit protein machines is emerging as a cornerstone of modern biology, and molecular-level study of these machines in most cases will require recombinant production. Here, we present and validate a strategy to rapidly produce, permutate, and posttranslationally modify large, eukaryotic multiprotein complexes by using DNA recombination in a process that is fully automatable. Parallel production of 12 protein complex variants within a period of weeks resulted in specimens of sufficient quantity and homogeneity for structural biology applications.

KW - Animals

KW - Baculoviridae

KW - Cell Line

KW - Escherichia coli

KW - Genetic Vectors

KW - Humans

KW - Multiprotein Complexes

KW - Spodoptera

U2 - 10.1016/j.str.2007.01.016

DO - 10.1016/j.str.2007.01.016

M3 - Article (Academic Journal)

C2 - 17355863

SN - 0969-2126

VL - 15

SP - 275

EP - 279

JO - Structure

JF - Structure

IS - 3

ER -

Multiprotein expression strategy for structural biology of eukaryotic complexes

Abstract

Keywords

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