Synthesis of the basic-helix-loop-helix region of the immunoglobulin enhancer binding protein E47 and evaluation of its structural and DNA binding properties

P. Bishop, C. Jones, Indraneel Ghosh, J. Chmielewski

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The basic-helix-loop-helix (bHLH) region of the immunoglobulin enhancer binding protein E47 (IEB E47) was prepared in high yield by a solid-phase peptide synthesis methodology. Size-exclusion chromatography, sedimentation equilibrium and cross-linking data showed that the synthetic bHLH protein, 1, was dimeric, and higher-order aggregates of trimer and tetramer were also observed. The circular dichroism spectrum of 1 showed a high helical content, which increased upon addition of DNA. containing the κE2 sequence. Gel mobility shift experiments showed that protein 1 bound sequence specifically to the κE2 sequence with a binding constant of 10-10 M2, and had an affinity for other E box sequences as well. Comparisons between the co-crystal structure of IEB E47 with DNA and structural studies in solution showed lower helical contents in solution as would have been predicted from the crystal structure.

Original languageEnglish (US)
Pages (from-to)149-154
Number of pages6
JournalInternational Journal of Peptide and Protein Research
Volume46
Issue number2
StatePublished - 1995
Externally publishedYes

Fingerprint

NF-kappa B
Crystal structure
E-Box Elements
Solid-Phase Synthesis Techniques
Size exclusion chromatography
DNA
Circular Dichroism
Sedimentation
Gel Chromatography
Proteins
Gels
Peptides
Experiments

Keywords

  • DNa binding
  • Helix-loop-helix
  • Solid-phase synthesis

ASJC Scopus subject areas

  • Biochemistry

Cite this

@article{04079968bdc64f0b9d215bd172c4c709,
title = "Synthesis of the basic-helix-loop-helix region of the immunoglobulin enhancer binding protein E47 and evaluation of its structural and DNA binding properties",
abstract = "The basic-helix-loop-helix (bHLH) region of the immunoglobulin enhancer binding protein E47 (IEB E47) was prepared in high yield by a solid-phase peptide synthesis methodology. Size-exclusion chromatography, sedimentation equilibrium and cross-linking data showed that the synthetic bHLH protein, 1, was dimeric, and higher-order aggregates of trimer and tetramer were also observed. The circular dichroism spectrum of 1 showed a high helical content, which increased upon addition of DNA. containing the κE2 sequence. Gel mobility shift experiments showed that protein 1 bound sequence specifically to the κE2 sequence with a binding constant of 10-10 M2, and had an affinity for other E box sequences as well. Comparisons between the co-crystal structure of IEB E47 with DNA and structural studies in solution showed lower helical contents in solution as would have been predicted from the crystal structure.",
keywords = "DNa binding, Helix-loop-helix, Solid-phase synthesis",
author = "P. Bishop and C. Jones and Indraneel Ghosh and J. Chmielewski",
year = "1995",
language = "English (US)",
volume = "46",
pages = "149--154",
journal = "International Journal of Peptide and Protein Research",
issn = "0367-8377",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - Synthesis of the basic-helix-loop-helix region of the immunoglobulin enhancer binding protein E47 and evaluation of its structural and DNA binding properties

AU - Bishop, P.

AU - Jones, C.

AU - Ghosh, Indraneel

AU - Chmielewski, J.

PY - 1995

Y1 - 1995

N2 - The basic-helix-loop-helix (bHLH) region of the immunoglobulin enhancer binding protein E47 (IEB E47) was prepared in high yield by a solid-phase peptide synthesis methodology. Size-exclusion chromatography, sedimentation equilibrium and cross-linking data showed that the synthetic bHLH protein, 1, was dimeric, and higher-order aggregates of trimer and tetramer were also observed. The circular dichroism spectrum of 1 showed a high helical content, which increased upon addition of DNA. containing the κE2 sequence. Gel mobility shift experiments showed that protein 1 bound sequence specifically to the κE2 sequence with a binding constant of 10-10 M2, and had an affinity for other E box sequences as well. Comparisons between the co-crystal structure of IEB E47 with DNA and structural studies in solution showed lower helical contents in solution as would have been predicted from the crystal structure.

AB - The basic-helix-loop-helix (bHLH) region of the immunoglobulin enhancer binding protein E47 (IEB E47) was prepared in high yield by a solid-phase peptide synthesis methodology. Size-exclusion chromatography, sedimentation equilibrium and cross-linking data showed that the synthetic bHLH protein, 1, was dimeric, and higher-order aggregates of trimer and tetramer were also observed. The circular dichroism spectrum of 1 showed a high helical content, which increased upon addition of DNA. containing the κE2 sequence. Gel mobility shift experiments showed that protein 1 bound sequence specifically to the κE2 sequence with a binding constant of 10-10 M2, and had an affinity for other E box sequences as well. Comparisons between the co-crystal structure of IEB E47 with DNA and structural studies in solution showed lower helical contents in solution as would have been predicted from the crystal structure.

KW - DNa binding

KW - Helix-loop-helix

KW - Solid-phase synthesis

UR - http://www.scopus.com/inward/record.url?scp=0029163156&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029163156&partnerID=8YFLogxK

M3 - Article

C2 - 8567169

AN - SCOPUS:0029163156

VL - 46

SP - 149

EP - 154

JO - International Journal of Peptide and Protein Research

JF - International Journal of Peptide and Protein Research

SN - 0367-8377

IS - 2

ER -