Overview of the Pipeline for Structural and Functional Characterization of Macrophage Proteins at the University of Queensland

Weining Meng; Jade K. Forwood; Gregor Guncar; Gautier Robin; Nathan P. Cowieson; Pawel Listwan; Dmitri Mouradov; Gordon King; Ian L. Ross; Jodie Robinson; Munish Puri; Justine M. Hill; Stuart Kellie; Thomas Huber; David A. Hume; Jennifer L. Martin; Bostjan Kobe

doi:10.1007/978-1-60327-058-8_38

Overview of the Pipeline for Structural and Functional Characterization of Macrophage Proteins at the University of Queensland

Weining Meng, Jade K. Forwood, Gregor Guncar, Gautier Robin, Nathan P. Cowieson, Pawel Listwan, Dmitri Mouradov, Gordon King, Ian L. Ross, Jodie Robinson, Munish Puri, Justine M. Hill, Stuart Kellie, Thomas Huber, David A. Hume, Jennifer L. Martin, Bostjan Kobe

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

2 Citations (Scopus)

Abstract

This chapter describes the methodology adopted in a project aimed at structural and functional characterization of proteins that potentially play an important role in mammalian macrophages. The methodology that underpins this project is applicable to both small research groups and larger structural genomics consortia. Gene products with putative roles in macrophage function are identified using gene expression information obtained via DNA microarray technology. Specific targets for structural and functional characterization are then selected based on a set of criteria aimed at maximizing insight into function. The target proteins are cloned using a modification of Gateway® cloning technology, expressed with hexa-histidine tags in E. coli, and purified to homogeneity using a combination of affinity and size exclusion chromatography. Purified proteins are finally subjected to crystallization trials and/or NMR-based screening to identify candidates for structure determination. Where crystallography and NMR approaches are unsuccessful, chemical cross-linking is employed to obtain structural information. This resulting structural information is used to guide cell biology experiments to further investigate the cellular and molecular function of the targets in macrophage biology. Jointly, the data sheds light on the molecular and cellular functions of macrophage proteins.

Original language	English
Title of host publication	Structural Proteomics
Subtitle of host publication	High-Throughput Methods
Editors	Bostjan Kobe, Mitchell Guss, Thomas Huber
Place of Publication	New Jersey
Publisher	Humana Press
Chapter	38
Pages	577-587
Number of pages	11
ISBN (Electronic)	9781603270588
ISBN (Print)	9781588298096
DOIs	https://doi.org/10.1007/978-1-60327-058-8_38
Publication status	Published - 2008
Externally published	Yes

Publication series

Name	Methods in Molecular Biology
ISSN (Print)	1064-3745

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Meng, W., Forwood, J. K., Guncar, G., Robin, G., Cowieson, N. P., Listwan, P., Mouradov, D., King, G., Ross, I. L., Robinson, J., Puri, M., Hill, J. M., Kellie, S., Huber, T., Hume, D. A., Martin, J. L., & Kobe, B. (2008). Overview of the Pipeline for Structural and Functional Characterization of Macrophage Proteins at the University of Queensland. In B. Kobe, M. Guss, & T. Huber (Eds.), Structural Proteomics: High-Throughput Methods (pp. 577-587). (Methods in Molecular Biology). Humana Press. https://doi.org/10.1007/978-1-60327-058-8_38

@inbook{8c40dc7ffdc844a8bc6315d49975ea4d,

title = "Overview of the Pipeline for Structural and Functional Characterization of Macrophage Proteins at the University of Queensland",

abstract = "This chapter describes the methodology adopted in a project aimed at structural and functional characterization of proteins that potentially play an important role in mammalian macrophages. The methodology that underpins this project is applicable to both small research groups and larger structural genomics consortia. Gene products with putative roles in macrophage function are identified using gene expression information obtained via DNA microarray technology. Specific targets for structural and functional characterization are then selected based on a set of criteria aimed at maximizing insight into function. The target proteins are cloned using a modification of Gateway{\textregistered} cloning technology, expressed with hexa-histidine tags in E. coli, and purified to homogeneity using a combination of affinity and size exclusion chromatography. Purified proteins are finally subjected to crystallization trials and/or NMR-based screening to identify candidates for structure determination. Where crystallography and NMR approaches are unsuccessful, chemical cross-linking is employed to obtain structural information. This resulting structural information is used to guide cell biology experiments to further investigate the cellular and molecular function of the targets in macrophage biology. Jointly, the data sheds light on the molecular and cellular functions of macrophage proteins.",

author = "Weining Meng and Forwood, {Jade K.} and Gregor Guncar and Gautier Robin and Cowieson, {Nathan P.} and Pawel Listwan and Dmitri Mouradov and Gordon King and Ross, {Ian L.} and Jodie Robinson and Munish Puri and Hill, {Justine M.} and Stuart Kellie and Thomas Huber and Hume, {David A.} and Martin, {Jennifer L.} and Bostjan Kobe",

year = "2008",

doi = "10.1007/978-1-60327-058-8_38",

language = "English",

isbn = "9781588298096",

series = "Methods in Molecular Biology",

publisher = "Humana Press",

pages = "577--587",

editor = "Kobe, {Bostjan } and Mitchell Guss and Thomas Huber",

booktitle = "Structural Proteomics",

address = "United States",

}

Meng, W, Forwood, JK, Guncar, G, Robin, G, Cowieson, NP, Listwan, P, Mouradov, D, King, G, Ross, IL, Robinson, J, Puri, M, Hill, JM, Kellie, S, Huber, T, Hume, DA, Martin, JL & Kobe, B 2008, Overview of the Pipeline for Structural and Functional Characterization of Macrophage Proteins at the University of Queensland. in B Kobe, M Guss & T Huber (eds), Structural Proteomics: High-Throughput Methods. Methods in Molecular Biology, Humana Press, New Jersey, pp. 577-587. https://doi.org/10.1007/978-1-60327-058-8_38

Overview of the Pipeline for Structural and Functional Characterization of Macrophage Proteins at the University of Queensland. / Meng, Weining; Forwood, Jade K.; Guncar, Gregor et al.
Structural Proteomics: High-Throughput Methods. ed. / Bostjan Kobe; Mitchell Guss; Thomas Huber. New Jersey: Humana Press, 2008. p. 577-587 (Methods in Molecular Biology).

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

TY - CHAP

T1 - Overview of the Pipeline for Structural and Functional Characterization of Macrophage Proteins at the University of Queensland

AU - Meng, Weining

AU - Forwood, Jade K.

AU - Guncar, Gregor

AU - Robin, Gautier

AU - Cowieson, Nathan P.

AU - Listwan, Pawel

AU - Mouradov, Dmitri

AU - King, Gordon

AU - Ross, Ian L.

AU - Robinson, Jodie

AU - Puri, Munish

AU - Hill, Justine M.

AU - Kellie, Stuart

AU - Huber, Thomas

AU - Hume, David A.

AU - Martin, Jennifer L.

AU - Kobe, Bostjan

PY - 2008

Y1 - 2008

N2 - This chapter describes the methodology adopted in a project aimed at structural and functional characterization of proteins that potentially play an important role in mammalian macrophages. The methodology that underpins this project is applicable to both small research groups and larger structural genomics consortia. Gene products with putative roles in macrophage function are identified using gene expression information obtained via DNA microarray technology. Specific targets for structural and functional characterization are then selected based on a set of criteria aimed at maximizing insight into function. The target proteins are cloned using a modification of Gateway® cloning technology, expressed with hexa-histidine tags in E. coli, and purified to homogeneity using a combination of affinity and size exclusion chromatography. Purified proteins are finally subjected to crystallization trials and/or NMR-based screening to identify candidates for structure determination. Where crystallography and NMR approaches are unsuccessful, chemical cross-linking is employed to obtain structural information. This resulting structural information is used to guide cell biology experiments to further investigate the cellular and molecular function of the targets in macrophage biology. Jointly, the data sheds light on the molecular and cellular functions of macrophage proteins.

AB - This chapter describes the methodology adopted in a project aimed at structural and functional characterization of proteins that potentially play an important role in mammalian macrophages. The methodology that underpins this project is applicable to both small research groups and larger structural genomics consortia. Gene products with putative roles in macrophage function are identified using gene expression information obtained via DNA microarray technology. Specific targets for structural and functional characterization are then selected based on a set of criteria aimed at maximizing insight into function. The target proteins are cloned using a modification of Gateway® cloning technology, expressed with hexa-histidine tags in E. coli, and purified to homogeneity using a combination of affinity and size exclusion chromatography. Purified proteins are finally subjected to crystallization trials and/or NMR-based screening to identify candidates for structure determination. Where crystallography and NMR approaches are unsuccessful, chemical cross-linking is employed to obtain structural information. This resulting structural information is used to guide cell biology experiments to further investigate the cellular and molecular function of the targets in macrophage biology. Jointly, the data sheds light on the molecular and cellular functions of macrophage proteins.

U2 - 10.1007/978-1-60327-058-8_38

DO - 10.1007/978-1-60327-058-8_38

M3 - Chapter

SN - 9781588298096

T3 - Methods in Molecular Biology

SP - 577

EP - 587

BT - Structural Proteomics

A2 - Kobe, Bostjan

A2 - Guss, Mitchell

A2 - Huber, Thomas

PB - Humana Press

CY - New Jersey

ER -

Meng W, Forwood JK, Guncar G, Robin G, Cowieson NP, Listwan P et al. Overview of the Pipeline for Structural and Functional Characterization of Macrophage Proteins at the University of Queensland. In Kobe B, Guss M, Huber T, editors, Structural Proteomics: High-Throughput Methods. New Jersey: Humana Press. 2008. p. 577-587. (Methods in Molecular Biology). doi: 10.1007/978-1-60327-058-8_38

Overview of the Pipeline for Structural and Functional Characterization of Macrophage Proteins at the University of Queensland

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