Unfolding and Refolding of Juvenile Hormone Binding Protein

Juvenile hormone (JH) regulates insect development. JH present in the hemolymph is bound to a specific glycoprotein, juvenile hormone binding protein (JHBP), which serves as a carrier to deploy the hormone to target tissues. In this report structural changes of JHBP from Galleria mellonella induced...

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Main Authors: Dobryszycki, Piotr, Kołodziejczyk, Robert, Krowarsch, Daniel, Gapiński, Jacek, Ożyhar, Andrzej, Kochman, Marian
Formato: Artigo
Idioma:English
Publicado em: Biophysical Society 2004
Assuntos:
Proteins
Acesso em linha:https://ncbi.nlm.nih.gov/pmc/articles/PMC1303906/
https://ncbi.nlm.nih.gov/pubmed/14747348
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spelling pubmed-13039062006-01-17 Unfolding and Refolding of Juvenile Hormone Binding Protein Dobryszycki, Piotr Kołodziejczyk, Robert Krowarsch, Daniel Gapiński, Jacek Ożyhar, Andrzej Kochman, Marian Biophys J Proteins Juvenile hormone (JH) regulates insect development. JH present in the hemolymph is bound to a specific glycoprotein, juvenile hormone binding protein (JHBP), which serves as a carrier to deploy the hormone to target tissues. In this report structural changes of JHBP from Galleria mellonella induced by guanidine hydrochloride have been investigated by a combination of size-exclusion chromatography, protein activity measurements, and spectroscopic methods. Molecules of JHBP change their conformation from a native state via two unstable intermediates to a denatured state. The first intermediate appears in a compact state, because it slightly changes its molecular size and preserves most of the JHBP secondary structure of the native state. Although the second intermediate also preserves a substantial part of the secondary structure, it undergoes a change into a noncompact state changing its Stokes radius from ∼30 to 39 Å. Refolding experiments showed that JHBP molecules recover their full protein structure, as judged from the CD spectrum, fluorescence experiments, and JH binding activity measurements. The free energy of unfolding in the absence of the denaturant, ΔG(D–N), is calculated to be 4.1 kcal mol(−1). Biophysical Society 2004-02 /pmc/articles/PMC1303906/ /pubmed/14747348 Text en Copyright © 2004, Biophysical Society
institution US National Library of Medicine
collection PubMed Central
language English
format Article
topic Proteins
spellingShingle Proteins
Dobryszycki, Piotr
Kołodziejczyk, Robert
Krowarsch, Daniel
Gapiński, Jacek
Ożyhar, Andrzej
Kochman, Marian
Unfolding and Refolding of Juvenile Hormone Binding Protein
description Juvenile hormone (JH) regulates insect development. JH present in the hemolymph is bound to a specific glycoprotein, juvenile hormone binding protein (JHBP), which serves as a carrier to deploy the hormone to target tissues. In this report structural changes of JHBP from Galleria mellonella induced by guanidine hydrochloride have been investigated by a combination of size-exclusion chromatography, protein activity measurements, and spectroscopic methods. Molecules of JHBP change their conformation from a native state via two unstable intermediates to a denatured state. The first intermediate appears in a compact state, because it slightly changes its molecular size and preserves most of the JHBP secondary structure of the native state. Although the second intermediate also preserves a substantial part of the secondary structure, it undergoes a change into a noncompact state changing its Stokes radius from ∼30 to 39 Å. Refolding experiments showed that JHBP molecules recover their full protein structure, as judged from the CD spectrum, fluorescence experiments, and JH binding activity measurements. The free energy of unfolding in the absence of the denaturant, ΔG(D–N), is calculated to be 4.1 kcal mol(−1).
author Dobryszycki, Piotr
Kołodziejczyk, Robert
Krowarsch, Daniel
Gapiński, Jacek
Ożyhar, Andrzej
Kochman, Marian
author_facet Dobryszycki, Piotr
Kołodziejczyk, Robert
Krowarsch, Daniel
Gapiński, Jacek
Ożyhar, Andrzej
Kochman, Marian
author_sort Dobryszycki, Piotr
title Unfolding and Refolding of Juvenile Hormone Binding Protein
title_short Unfolding and Refolding of Juvenile Hormone Binding Protein
title_full Unfolding and Refolding of Juvenile Hormone Binding Protein
title_fullStr Unfolding and Refolding of Juvenile Hormone Binding Protein
title_full_unstemmed Unfolding and Refolding of Juvenile Hormone Binding Protein
title_sort unfolding and refolding of juvenile hormone binding protein
publisher Biophysical Society
publisher_facet Biophysical Society
publishDate 2004
url https://ncbi.nlm.nih.gov/pmc/articles/PMC1303906/
https://ncbi.nlm.nih.gov/pubmed/14747348
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