Protein microarrays to detect protein-protein interactions using red and green fluorescent proteins

Thomas Kukar, Sarah Eckenrode, Yunrong Gu, Wei Lian, Mike Megginson, Jin-Xiong She, Donghai Wu

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

Proteomics, the study of protein function on a global scale, will play an important role in furthering our understanding of gene functions, complex biological pathways, and discovery of novel drug targets. A number of techniques have been developed for proteomic studies to identify and analyze proteins, compare protein expression levels, and study protein-protein interactions. Recent developments have applied a DNA array-type approach to immobilize proteins on a surface for high-throughput analysis. Here we report the development and construction of protein chips using derivatized glass and nitrocellulose-coated slides and the employment of recombinant proteins fused with green and red fluorescent proteins for detection. Fluorescent signals were found to be proportional to the amount of arrayed proteins and could be readily detected with a conventional fluorescence slide scanner. This technique allows the investigation of protein-protein interactions without the need for additional labeling steps of probe proteins.

Original languageEnglish (US)
Pages (from-to)50-54
Number of pages5
JournalAnalytical Biochemistry
Volume306
Issue number1
DOIs
StatePublished - Jul 1 2002
Externally publishedYes

Fingerprint

Protein Array Analysis
Microarrays
Green Fluorescent Proteins
Proteins
Proteomics
red fluorescent protein
Collodion
Drug Discovery
Oligonucleotide Array Sequence Analysis
Recombinant Proteins
Glass
Labeling
Fluorescence
Genes
Throughput

Keywords

  • Fluorescent proteins
  • Green fluorescent protein
  • Microarrays
  • Protein interactions
  • Red fluorescent protein

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Protein microarrays to detect protein-protein interactions using red and green fluorescent proteins. / Kukar, Thomas; Eckenrode, Sarah; Gu, Yunrong; Lian, Wei; Megginson, Mike; She, Jin-Xiong; Wu, Donghai.

In: Analytical Biochemistry, Vol. 306, No. 1, 01.07.2002, p. 50-54.

Research output: Contribution to journalArticle

Kukar, Thomas ; Eckenrode, Sarah ; Gu, Yunrong ; Lian, Wei ; Megginson, Mike ; She, Jin-Xiong ; Wu, Donghai. / Protein microarrays to detect protein-protein interactions using red and green fluorescent proteins. In: Analytical Biochemistry. 2002 ; Vol. 306, No. 1. pp. 50-54.
@article{59fbd9ad782745cfa1dc15c5f40b00ad,
title = "Protein microarrays to detect protein-protein interactions using red and green fluorescent proteins",
abstract = "Proteomics, the study of protein function on a global scale, will play an important role in furthering our understanding of gene functions, complex biological pathways, and discovery of novel drug targets. A number of techniques have been developed for proteomic studies to identify and analyze proteins, compare protein expression levels, and study protein-protein interactions. Recent developments have applied a DNA array-type approach to immobilize proteins on a surface for high-throughput analysis. Here we report the development and construction of protein chips using derivatized glass and nitrocellulose-coated slides and the employment of recombinant proteins fused with green and red fluorescent proteins for detection. Fluorescent signals were found to be proportional to the amount of arrayed proteins and could be readily detected with a conventional fluorescence slide scanner. This technique allows the investigation of protein-protein interactions without the need for additional labeling steps of probe proteins.",
keywords = "Fluorescent proteins, Green fluorescent protein, Microarrays, Protein interactions, Red fluorescent protein",
author = "Thomas Kukar and Sarah Eckenrode and Yunrong Gu and Wei Lian and Mike Megginson and Jin-Xiong She and Donghai Wu",
year = "2002",
month = "7",
day = "1",
doi = "10.1006/abio.2002.5614",
language = "English (US)",
volume = "306",
pages = "50--54",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Protein microarrays to detect protein-protein interactions using red and green fluorescent proteins

AU - Kukar, Thomas

AU - Eckenrode, Sarah

AU - Gu, Yunrong

AU - Lian, Wei

AU - Megginson, Mike

AU - She, Jin-Xiong

AU - Wu, Donghai

PY - 2002/7/1

Y1 - 2002/7/1

N2 - Proteomics, the study of protein function on a global scale, will play an important role in furthering our understanding of gene functions, complex biological pathways, and discovery of novel drug targets. A number of techniques have been developed for proteomic studies to identify and analyze proteins, compare protein expression levels, and study protein-protein interactions. Recent developments have applied a DNA array-type approach to immobilize proteins on a surface for high-throughput analysis. Here we report the development and construction of protein chips using derivatized glass and nitrocellulose-coated slides and the employment of recombinant proteins fused with green and red fluorescent proteins for detection. Fluorescent signals were found to be proportional to the amount of arrayed proteins and could be readily detected with a conventional fluorescence slide scanner. This technique allows the investigation of protein-protein interactions without the need for additional labeling steps of probe proteins.

AB - Proteomics, the study of protein function on a global scale, will play an important role in furthering our understanding of gene functions, complex biological pathways, and discovery of novel drug targets. A number of techniques have been developed for proteomic studies to identify and analyze proteins, compare protein expression levels, and study protein-protein interactions. Recent developments have applied a DNA array-type approach to immobilize proteins on a surface for high-throughput analysis. Here we report the development and construction of protein chips using derivatized glass and nitrocellulose-coated slides and the employment of recombinant proteins fused with green and red fluorescent proteins for detection. Fluorescent signals were found to be proportional to the amount of arrayed proteins and could be readily detected with a conventional fluorescence slide scanner. This technique allows the investigation of protein-protein interactions without the need for additional labeling steps of probe proteins.

KW - Fluorescent proteins

KW - Green fluorescent protein

KW - Microarrays

KW - Protein interactions

KW - Red fluorescent protein

UR - http://www.scopus.com/inward/record.url?scp=0036629196&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036629196&partnerID=8YFLogxK

U2 - 10.1006/abio.2002.5614

DO - 10.1006/abio.2002.5614

M3 - Article

VL - 306

SP - 50

EP - 54

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 1

ER -