An enhanced protein-protein interaction based on enzymatic complex through replacement of the recognition site

Sang Duck Jeon, Su Jung Kim, Sung Hyun Park, Gi Wook Choi, Sung Ok Han

Research output: Contribution to journalArticle

Abstract

Clostridium cellulovorans, produce multi-enzymatic complexes known as cellulosomes, which assemble via the interaction of a dockerin module in the cellulosomal subunit with one of the several cohesin modules in the scaffolding protein, to degrade the plant cell wall polymer. An enhanced cohesin-dockerin interaction was demonstrated by modified certain cellulosomal enzymes with altered amino acid residues at the crucial binding site, 11th and 12th positions in dockerin module. In fluorescence intensity analyses using the cellulosome-based biomarker system, the modified cellulosomal enzymes (EngE SL to AI and EngH SM to AI) showed an increased intensity (1.4- to 2.2-fold) compared with the wild-type proteins. Conversely, modified ExgS (AI to SM) exhibited a reduced intensity (0.6- to 0.7-fold) compared with the wild type. In enzyme-linked and competitive enzyme-linked interaction assays, the some modified protein (EngE SL to AI and EngH SM to AI) showed their increased binding affinity toward the cohesins (Coh2 and Coh9). Surface plasmon resonance analysis quantitatively demonstrated the binding affinity of these two modified proteins toward cohesins showed similar or higher affinity comparing with its with wild type proteins. These results suggest the replacement of amino acid residues in the certain recognition site significantly affects the binding affinity of the cohesin-dockerin interaction.

Original languageEnglish
Pages (from-to)1-6
Number of pages6
JournalInternational Journal of Biological Macromolecules
Volume75
DOIs
Publication statusPublished - 2015 Apr 1

Fingerprint

Cellulosomes
Enzymes
Proteins
Clostridium cellulovorans
Amino Acids
Clostridium
Surface Plasmon Resonance
Plant Cells
Surface plasmon resonance
Biomarkers
Cell Wall
Assays
Polymers
Fluorescence
Binding Sites
cohesins

Keywords

  • Cellulosome
  • Clostridium cellulovorans
  • Cohesin-dockerin interaction

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Cite this

An enhanced protein-protein interaction based on enzymatic complex through replacement of the recognition site. / Jeon, Sang Duck; Kim, Su Jung; Park, Sung Hyun; Choi, Gi Wook; Han, Sung Ok.

In: International Journal of Biological Macromolecules, Vol. 75, 01.04.2015, p. 1-6.

Research output: Contribution to journalArticle

@article{b345dec124794200a2f749ce61a70b9e,
title = "An enhanced protein-protein interaction based on enzymatic complex through replacement of the recognition site",
abstract = "Clostridium cellulovorans, produce multi-enzymatic complexes known as cellulosomes, which assemble via the interaction of a dockerin module in the cellulosomal subunit with one of the several cohesin modules in the scaffolding protein, to degrade the plant cell wall polymer. An enhanced cohesin-dockerin interaction was demonstrated by modified certain cellulosomal enzymes with altered amino acid residues at the crucial binding site, 11th and 12th positions in dockerin module. In fluorescence intensity analyses using the cellulosome-based biomarker system, the modified cellulosomal enzymes (EngE SL to AI and EngH SM to AI) showed an increased intensity (1.4- to 2.2-fold) compared with the wild-type proteins. Conversely, modified ExgS (AI to SM) exhibited a reduced intensity (0.6- to 0.7-fold) compared with the wild type. In enzyme-linked and competitive enzyme-linked interaction assays, the some modified protein (EngE SL to AI and EngH SM to AI) showed their increased binding affinity toward the cohesins (Coh2 and Coh9). Surface plasmon resonance analysis quantitatively demonstrated the binding affinity of these two modified proteins toward cohesins showed similar or higher affinity comparing with its with wild type proteins. These results suggest the replacement of amino acid residues in the certain recognition site significantly affects the binding affinity of the cohesin-dockerin interaction.",
keywords = "Cellulosome, Clostridium cellulovorans, Cohesin-dockerin interaction",
author = "Jeon, {Sang Duck} and Kim, {Su Jung} and Park, {Sung Hyun} and Choi, {Gi Wook} and Han, {Sung Ok}",
year = "2015",
month = "4",
day = "1",
doi = "10.1016/j.ijbiomac.2015.01.018",
language = "English",
volume = "75",
pages = "1--6",
journal = "International Journal of Biological Macromolecules",
issn = "0141-8130",
publisher = "Elsevier",

}

TY - JOUR

T1 - An enhanced protein-protein interaction based on enzymatic complex through replacement of the recognition site

AU - Jeon, Sang Duck

AU - Kim, Su Jung

AU - Park, Sung Hyun

AU - Choi, Gi Wook

AU - Han, Sung Ok

PY - 2015/4/1

Y1 - 2015/4/1

N2 - Clostridium cellulovorans, produce multi-enzymatic complexes known as cellulosomes, which assemble via the interaction of a dockerin module in the cellulosomal subunit with one of the several cohesin modules in the scaffolding protein, to degrade the plant cell wall polymer. An enhanced cohesin-dockerin interaction was demonstrated by modified certain cellulosomal enzymes with altered amino acid residues at the crucial binding site, 11th and 12th positions in dockerin module. In fluorescence intensity analyses using the cellulosome-based biomarker system, the modified cellulosomal enzymes (EngE SL to AI and EngH SM to AI) showed an increased intensity (1.4- to 2.2-fold) compared with the wild-type proteins. Conversely, modified ExgS (AI to SM) exhibited a reduced intensity (0.6- to 0.7-fold) compared with the wild type. In enzyme-linked and competitive enzyme-linked interaction assays, the some modified protein (EngE SL to AI and EngH SM to AI) showed their increased binding affinity toward the cohesins (Coh2 and Coh9). Surface plasmon resonance analysis quantitatively demonstrated the binding affinity of these two modified proteins toward cohesins showed similar or higher affinity comparing with its with wild type proteins. These results suggest the replacement of amino acid residues in the certain recognition site significantly affects the binding affinity of the cohesin-dockerin interaction.

AB - Clostridium cellulovorans, produce multi-enzymatic complexes known as cellulosomes, which assemble via the interaction of a dockerin module in the cellulosomal subunit with one of the several cohesin modules in the scaffolding protein, to degrade the plant cell wall polymer. An enhanced cohesin-dockerin interaction was demonstrated by modified certain cellulosomal enzymes with altered amino acid residues at the crucial binding site, 11th and 12th positions in dockerin module. In fluorescence intensity analyses using the cellulosome-based biomarker system, the modified cellulosomal enzymes (EngE SL to AI and EngH SM to AI) showed an increased intensity (1.4- to 2.2-fold) compared with the wild-type proteins. Conversely, modified ExgS (AI to SM) exhibited a reduced intensity (0.6- to 0.7-fold) compared with the wild type. In enzyme-linked and competitive enzyme-linked interaction assays, the some modified protein (EngE SL to AI and EngH SM to AI) showed their increased binding affinity toward the cohesins (Coh2 and Coh9). Surface plasmon resonance analysis quantitatively demonstrated the binding affinity of these two modified proteins toward cohesins showed similar or higher affinity comparing with its with wild type proteins. These results suggest the replacement of amino acid residues in the certain recognition site significantly affects the binding affinity of the cohesin-dockerin interaction.

KW - Cellulosome

KW - Clostridium cellulovorans

KW - Cohesin-dockerin interaction

UR - http://www.scopus.com/inward/record.url?scp=84921843009&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84921843009&partnerID=8YFLogxK

U2 - 10.1016/j.ijbiomac.2015.01.018

DO - 10.1016/j.ijbiomac.2015.01.018

M3 - Article

C2 - 25603141

AN - SCOPUS:84921843009

VL - 75

SP - 1

EP - 6

JO - International Journal of Biological Macromolecules

JF - International Journal of Biological Macromolecules

SN - 0141-8130

ER -