Identification of HMGB1-Binding Components Using Affinity Column Chromatography

Ari Rouhiainen, Helena Tukiainen, Pia Siljander

Forskningsoutput: Kapitel i bok/rapport/konferenshandlingKapitelVetenskaplig

Sammanfattning

High Mobility Group B1 (HMGB1) is a 30 kDa protein widely expressed in mammalian
cells. HMGB1 has a high content of charged amino acids and has a bipolar structure
consisting of two highly positive amino terminal HMG-box domains and an acidic carboxy
terminal tail. HMGB1 has nuclear functions regulating chromatin structure and gene
expression and extracellular functions regulating immune response and cell motility.
Biochemical and cell biological studies have revealed that HMGB1 binds to various kinds
of biomolecules and these interactions are crucial for determining the in vivo functions of
HMGB1. Albeit several different biochemical methods have been used to detect HMGB1-
binding components, HMGB1-affinity column chromatography has rarely been applied in
such studies. Here, we describe an affinity chromatography method that we have applied
to isolation and identification of HMGB1-binding molecules from different cell types.
Biomolecules recovered with HMGB1-affinity chromatography include proinflammatory
bacterial DNA and glioblastoma cell histones H1 and H3 which all have previously been
reported as HMGB1-binding molecules by other methods. Furthermore, an entirely new
HMGB1-binding protein, Multimerin-1 containing complex, was identified from platelet
lysates by HMGB1-affinity chromatography. Endogenous Multimerin-1 and HMGB1 were
shown to associate on the surface of endothelial cells and activated platelets, and endogenous
Multimerin-1 also regulated the release of HMGB1 from activated platelets. In
conclusion, HMGB1-affinity chromatography can be used to isolate and characterize novel
HMGB1-binding partners from a variety of cellular sources. Such new interactions reveal
further complexity in the multi-faceted biology of the HMGB1.
Originalspråkengelska
Titel på gästpublikationProtein Engineering - Technology and Application
FörlagINTECHopen
Utgivningsdatum31 maj 2013
Sidor168-187
ISBN (tryckt)978-953-51-1138-2
DOI
StatusPublicerad - 31 maj 2013
MoE-publikationstypB2 Del av bok eller annan forskningsbok

Vetenskapsgrenar

  • 1182 Biokemi, cell- och molekylärbiologi

Citera det här

Rouhiainen, A., Tukiainen, H., & Siljander, P. (2013). Identification of HMGB1-Binding Components Using Affinity Column Chromatography. I Protein Engineering - Technology and Application (s. 168-187). INTECHopen. https://doi.org/10.5772/56336
Rouhiainen, Ari ; Tukiainen, Helena ; Siljander, Pia. / Identification of HMGB1-Binding Components Using Affinity Column Chromatography. Protein Engineering - Technology and Application. INTECHopen, 2013. s. 168-187
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title = "Identification of HMGB1-Binding Components Using Affinity Column Chromatography",
abstract = "High Mobility Group B1 (HMGB1) is a 30 kDa protein widely expressed in mammaliancells. HMGB1 has a high content of charged amino acids and has a bipolar structureconsisting of two highly positive amino terminal HMG-box domains and an acidic carboxyterminal tail. HMGB1 has nuclear functions regulating chromatin structure and geneexpression and extracellular functions regulating immune response and cell motility.Biochemical and cell biological studies have revealed that HMGB1 binds to various kindsof biomolecules and these interactions are crucial for determining the in vivo functions ofHMGB1. Albeit several different biochemical methods have been used to detect HMGB1-binding components, HMGB1-affinity column chromatography has rarely been applied insuch studies. Here, we describe an affinity chromatography method that we have appliedto isolation and identification of HMGB1-binding molecules from different cell types.Biomolecules recovered with HMGB1-affinity chromatography include proinflammatorybacterial DNA and glioblastoma cell histones H1 and H3 which all have previously beenreported as HMGB1-binding molecules by other methods. Furthermore, an entirely newHMGB1-binding protein, Multimerin-1 containing complex, was identified from plateletlysates by HMGB1-affinity chromatography. Endogenous Multimerin-1 and HMGB1 wereshown to associate on the surface of endothelial cells and activated platelets, and endogenousMultimerin-1 also regulated the release of HMGB1 from activated platelets. Inconclusion, HMGB1-affinity chromatography can be used to isolate and characterize novelHMGB1-binding partners from a variety of cellular sources. Such new interactions revealfurther complexity in the multi-faceted biology of the HMGB1.",
keywords = "1182 Biochemistry, cell and molecular biology, HMGB1, Multimerin-1 , Platelet, Endothelium, Coagulation",
author = "Ari Rouhiainen and Helena Tukiainen and Pia Siljander",
year = "2013",
month = "5",
day = "31",
doi = "10.5772/56336",
language = "English",
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Rouhiainen, A, Tukiainen, H & Siljander, P 2013, Identification of HMGB1-Binding Components Using Affinity Column Chromatography. i Protein Engineering - Technology and Application. INTECHopen, s. 168-187. https://doi.org/10.5772/56336

Identification of HMGB1-Binding Components Using Affinity Column Chromatography. / Rouhiainen, Ari; Tukiainen, Helena; Siljander, Pia.

Protein Engineering - Technology and Application. INTECHopen, 2013. s. 168-187.

Forskningsoutput: Kapitel i bok/rapport/konferenshandlingKapitelVetenskaplig

TY - CHAP

T1 - Identification of HMGB1-Binding Components Using Affinity Column Chromatography

AU - Rouhiainen, Ari

AU - Tukiainen, Helena

AU - Siljander, Pia

PY - 2013/5/31

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N2 - High Mobility Group B1 (HMGB1) is a 30 kDa protein widely expressed in mammaliancells. HMGB1 has a high content of charged amino acids and has a bipolar structureconsisting of two highly positive amino terminal HMG-box domains and an acidic carboxyterminal tail. HMGB1 has nuclear functions regulating chromatin structure and geneexpression and extracellular functions regulating immune response and cell motility.Biochemical and cell biological studies have revealed that HMGB1 binds to various kindsof biomolecules and these interactions are crucial for determining the in vivo functions ofHMGB1. Albeit several different biochemical methods have been used to detect HMGB1-binding components, HMGB1-affinity column chromatography has rarely been applied insuch studies. Here, we describe an affinity chromatography method that we have appliedto isolation and identification of HMGB1-binding molecules from different cell types.Biomolecules recovered with HMGB1-affinity chromatography include proinflammatorybacterial DNA and glioblastoma cell histones H1 and H3 which all have previously beenreported as HMGB1-binding molecules by other methods. Furthermore, an entirely newHMGB1-binding protein, Multimerin-1 containing complex, was identified from plateletlysates by HMGB1-affinity chromatography. Endogenous Multimerin-1 and HMGB1 wereshown to associate on the surface of endothelial cells and activated platelets, and endogenousMultimerin-1 also regulated the release of HMGB1 from activated platelets. Inconclusion, HMGB1-affinity chromatography can be used to isolate and characterize novelHMGB1-binding partners from a variety of cellular sources. Such new interactions revealfurther complexity in the multi-faceted biology of the HMGB1.

AB - High Mobility Group B1 (HMGB1) is a 30 kDa protein widely expressed in mammaliancells. HMGB1 has a high content of charged amino acids and has a bipolar structureconsisting of two highly positive amino terminal HMG-box domains and an acidic carboxyterminal tail. HMGB1 has nuclear functions regulating chromatin structure and geneexpression and extracellular functions regulating immune response and cell motility.Biochemical and cell biological studies have revealed that HMGB1 binds to various kindsof biomolecules and these interactions are crucial for determining the in vivo functions ofHMGB1. Albeit several different biochemical methods have been used to detect HMGB1-binding components, HMGB1-affinity column chromatography has rarely been applied insuch studies. Here, we describe an affinity chromatography method that we have appliedto isolation and identification of HMGB1-binding molecules from different cell types.Biomolecules recovered with HMGB1-affinity chromatography include proinflammatorybacterial DNA and glioblastoma cell histones H1 and H3 which all have previously beenreported as HMGB1-binding molecules by other methods. Furthermore, an entirely newHMGB1-binding protein, Multimerin-1 containing complex, was identified from plateletlysates by HMGB1-affinity chromatography. Endogenous Multimerin-1 and HMGB1 wereshown to associate on the surface of endothelial cells and activated platelets, and endogenousMultimerin-1 also regulated the release of HMGB1 from activated platelets. Inconclusion, HMGB1-affinity chromatography can be used to isolate and characterize novelHMGB1-binding partners from a variety of cellular sources. Such new interactions revealfurther complexity in the multi-faceted biology of the HMGB1.

KW - 1182 Biochemistry, cell and molecular biology

KW - HMGB1

KW - Multimerin-1

KW - Platelet

KW - Endothelium

KW - Coagulation

U2 - 10.5772/56336

DO - 10.5772/56336

M3 - Chapter

SN - 978-953-51-1138-2

SP - 168

EP - 187

BT - Protein Engineering - Technology and Application

PB - INTECHopen

ER -

Rouhiainen A, Tukiainen H, Siljander P. Identification of HMGB1-Binding Components Using Affinity Column Chromatography. I Protein Engineering - Technology and Application. INTECHopen. 2013. s. 168-187 https://doi.org/10.5772/56336