Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein

Autores
Curto, Lucrecia María; Caramelo, Julio Javier; Delfino, Jose Maria
Año de publicación
2005
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Intestinal fatty acid binding protein (IFABP) is a 15 kDa intracellular lipid-binding protein exhibiting a ́-barrel fold that resembles a clamshell. The ́-barrel, which encloses the ligand binding cavity, consists of two perpendicular five-stranded ́-sheets with an intervening helix-turn-helix motif between strands A and B. Δ98Δ (fragment 29-126 of IFABP) was obtained either in its recombinant form or by limited proteolysis with clostripain. Despite lacking extensive stretches involved in the closure of the β-barrel, Δ98Δ remains soluble and stable in solution. Spectroscopic analyses by circular dichroism, ultraviolet absorption, and intrinsic fluorescence indicate that the fragment retains substantial β-sheet content and tertiary interactions. In particular, the environment around W82 is identical in both Δ98Δ and IFABP, a fact consistent with the conservation in the former of all the critical amino acid residues belonging to the hydrophobic core. In addition, the Stokes radius of Δ98Δ is similar to that of IFABP and 16% larger than that calculated from its molecular weight (11 kDa). The monomeric status of Δ98Δ was further confirmed by chemical cross-linking experiments. Although lacking 25% of the amino acids of the parent protein, in the presence of GdnHCl, Δ98Δ unfolds through a cooperative transition showing a midpoint at 0.90 M. Remarkably, it also preserves binding activity for fatty acids (K d = 5.1 μM for oleic acid and Kd = 0.72 μM for trans-parinaric acid), a fact that exerts a stabilizing effect on its structure. These cumulative evidences show that Δ98Δ adopts a monomeric state with a compact core and a loose periphery, being so far the smallest structure of its kind preserving binding function.
Fil: Curto, Lucrecia María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Caramelo, Julio Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Delfino, Jose Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Materia
Ifabp
Abridged Variant
Truncation
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/39256
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/39256
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding proteinCurto, Lucrecia MaríaCaramelo, Julio JavierDelfino, Jose MariaIfabpAbridged VariantTruncationhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Intestinal fatty acid binding protein (IFABP) is a 15 kDa intracellular lipid-binding protein exhibiting a ́-barrel fold that resembles a clamshell. The ́-barrel, which encloses the ligand binding cavity, consists of two perpendicular five-stranded ́-sheets with an intervening helix-turn-helix motif between strands A and B. Δ98Δ (fragment 29-126 of IFABP) was obtained either in its recombinant form or by limited proteolysis with clostripain. Despite lacking extensive stretches involved in the closure of the β-barrel, Δ98Δ remains soluble and stable in solution. Spectroscopic analyses by circular dichroism, ultraviolet absorption, and intrinsic fluorescence indicate that the fragment retains substantial β-sheet content and tertiary interactions. In particular, the environment around W82 is identical in both Δ98Δ and IFABP, a fact consistent with the conservation in the former of all the critical amino acid residues belonging to the hydrophobic core. In addition, the Stokes radius of Δ98Δ is similar to that of IFABP and 16% larger than that calculated from its molecular weight (11 kDa). The monomeric status of Δ98Δ was further confirmed by chemical cross-linking experiments. Although lacking 25% of the amino acids of the parent protein, in the presence of GdnHCl, Δ98Δ unfolds through a cooperative transition showing a midpoint at 0.90 M. Remarkably, it also preserves binding activity for fatty acids (K d = 5.1 μM for oleic acid and Kd = 0.72 μM for trans-parinaric acid), a fact that exerts a stabilizing effect on its structure. These cumulative evidences show that Δ98Δ adopts a monomeric state with a compact core and a loose periphery, being so far the smallest structure of its kind preserving binding function.Fil: Curto, Lucrecia María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Caramelo, Julio Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Delfino, Jose Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaAmerican Chemical Society2005-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/39256Curto, Lucrecia María; Caramelo, Julio Javier; Delfino, Jose Maria; Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein; American Chemical Society; Biochemistry; 44; 42; 10-2005; 13847-138570006-29601520-4995CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/abs/10.1021/bi051080sinfo:eu-repo/semantics/altIdentifier/doi/10.1021/bi051080sinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:00:48Zoai:ri.conicet.gov.ar:11336/39256instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:00:48.938CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein
title Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein
spellingShingle Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein
Curto, Lucrecia María
Ifabp
Abridged Variant
Truncation
title_short Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein
title_full Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein
title_fullStr Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein
title_full_unstemmed Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein
title_sort Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein
dc.creator.none.fl_str_mv Curto, Lucrecia María
Caramelo, Julio Javier
Delfino, Jose Maria
author Curto, Lucrecia María
author_facet Curto, Lucrecia María
Caramelo, Julio Javier
Delfino, Jose Maria
author_role author
author2 Caramelo, Julio Javier
Delfino, Jose Maria
author2_role author
author
dc.subject.none.fl_str_mv Ifabp
Abridged Variant
Truncation
topic Ifabp
Abridged Variant
Truncation
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Intestinal fatty acid binding protein (IFABP) is a 15 kDa intracellular lipid-binding protein exhibiting a ́-barrel fold that resembles a clamshell. The ́-barrel, which encloses the ligand binding cavity, consists of two perpendicular five-stranded ́-sheets with an intervening helix-turn-helix motif between strands A and B. Δ98Δ (fragment 29-126 of IFABP) was obtained either in its recombinant form or by limited proteolysis with clostripain. Despite lacking extensive stretches involved in the closure of the β-barrel, Δ98Δ remains soluble and stable in solution. Spectroscopic analyses by circular dichroism, ultraviolet absorption, and intrinsic fluorescence indicate that the fragment retains substantial β-sheet content and tertiary interactions. In particular, the environment around W82 is identical in both Δ98Δ and IFABP, a fact consistent with the conservation in the former of all the critical amino acid residues belonging to the hydrophobic core. In addition, the Stokes radius of Δ98Δ is similar to that of IFABP and 16% larger than that calculated from its molecular weight (11 kDa). The monomeric status of Δ98Δ was further confirmed by chemical cross-linking experiments. Although lacking 25% of the amino acids of the parent protein, in the presence of GdnHCl, Δ98Δ unfolds through a cooperative transition showing a midpoint at 0.90 M. Remarkably, it also preserves binding activity for fatty acids (K d = 5.1 μM for oleic acid and Kd = 0.72 μM for trans-parinaric acid), a fact that exerts a stabilizing effect on its structure. These cumulative evidences show that Δ98Δ adopts a monomeric state with a compact core and a loose periphery, being so far the smallest structure of its kind preserving binding function.
Fil: Curto, Lucrecia María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Caramelo, Julio Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina
Fil: Delfino, Jose Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
description Intestinal fatty acid binding protein (IFABP) is a 15 kDa intracellular lipid-binding protein exhibiting a ́-barrel fold that resembles a clamshell. The ́-barrel, which encloses the ligand binding cavity, consists of two perpendicular five-stranded ́-sheets with an intervening helix-turn-helix motif between strands A and B. Δ98Δ (fragment 29-126 of IFABP) was obtained either in its recombinant form or by limited proteolysis with clostripain. Despite lacking extensive stretches involved in the closure of the β-barrel, Δ98Δ remains soluble and stable in solution. Spectroscopic analyses by circular dichroism, ultraviolet absorption, and intrinsic fluorescence indicate that the fragment retains substantial β-sheet content and tertiary interactions. In particular, the environment around W82 is identical in both Δ98Δ and IFABP, a fact consistent with the conservation in the former of all the critical amino acid residues belonging to the hydrophobic core. In addition, the Stokes radius of Δ98Δ is similar to that of IFABP and 16% larger than that calculated from its molecular weight (11 kDa). The monomeric status of Δ98Δ was further confirmed by chemical cross-linking experiments. Although lacking 25% of the amino acids of the parent protein, in the presence of GdnHCl, Δ98Δ unfolds through a cooperative transition showing a midpoint at 0.90 M. Remarkably, it also preserves binding activity for fatty acids (K d = 5.1 μM for oleic acid and Kd = 0.72 μM for trans-parinaric acid), a fact that exerts a stabilizing effect on its structure. These cumulative evidences show that Δ98Δ adopts a monomeric state with a compact core and a loose periphery, being so far the smallest structure of its kind preserving binding function.
publishDate 2005
dc.date.none.fl_str_mv 2005-10
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/39256
Curto, Lucrecia María; Caramelo, Julio Javier; Delfino, Jose Maria; Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein; American Chemical Society; Biochemistry; 44; 42; 10-2005; 13847-13857
0006-2960
1520-4995
CONICET Digital
CONICET
url http://hdl.handle.net/11336/39256
identifier_str_mv Curto, Lucrecia María; Caramelo, Julio Javier; Delfino, Jose Maria; Δ98Δ, a functional all-β-sheet abridged form of intestinal fatty acid binding protein; American Chemical Society; Biochemistry; 44; 42; 10-2005; 13847-13857
0006-2960
1520-4995
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://pubs.acs.org/doi/abs/10.1021/bi051080s
info:eu-repo/semantics/altIdentifier/doi/10.1021/bi051080s
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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