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dc.contributor.author
Gomez, Gabriela Elena
dc.contributor.author
Cauerhff, Ana
dc.contributor.author
Craig, Patricio Oliver
dc.contributor.author
Goldbaum, Fernando Alberto
dc.contributor.author
Delfino, Jose Maria
dc.date.available
2018-03-16T19:40:20Z
dc.date.issued
2006-04
dc.identifier.citation
Gomez, Gabriela Elena; Cauerhff, Ana; Craig, Patricio Oliver; Goldbaum, Fernando Alberto; Delfino, Jose Maria; Exploring protein interfaces with a general photochemical reagent; Cold Spring Harbor Lab Press; Protein Science; 15; 4; 4-2006; 744-752
dc.identifier.issn
0961-8368
dc.identifier.uri
http://hdl.handle.net/11336/39137
dc.description.abstract
Protein folding, natural conformational changes, or interaction between partners involved in recognition phenomena brings about differences in the solvent-accessible surface area (SASA) of the polypeptide chain. This primary event can be monitored by the differential chemical reactivity of functional groups along the protein sequence. Diazirine (DZN), a photoreactive gas similar in size to water, generates methylene carbene (:CH2). The extreme chemical reactivity of this species allows the almost instantaneous and indiscriminate modification of its immediate molecular cage. 3H-DZN was successfully used in our laboratory for studying protein structure and folding. Here we address for the first time the usefulness of this probe to examine the area of interaction in protein-protein complexes. For this purpose we chose the complex formed between hen eggwhite lysozyme(HEWL) and themonoclonal antibody IgG1D1.3. :CH2 labeling of free HEWL or complexed with IgG1 D1.3 yields 2.76 and 2.32 mmol CH 2 per mole protein at 1 mM DZN concentration, respectively. This reduction (15%) becomes consistent with the expected decrement in the SASA of HEWL occurring upon complexation derived from crystallographic data (11%), in agreement with the known unspecific surface labeling reaction of :CH 2. Further comparative analysis at the level of tryptic peptides led to the identification of the sites involved in the interaction. Remarkably, those peptides implicated in the contact area show the highest differential labeling: H15GLDNYR21, G117TDVQAWIR 125, and G22YSLGNWVCAAK33. Thus, protein footprinting with DZN emerges as a feasible methodology useful for mapping contact regions of protein domains involved in macromolecular assemblies.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Cold Spring Harbor Lab Press
dc.rights
info:eu-repo/semantics/restrictedAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Diazirine
dc.subject
Epitope Mapping
dc.subject
Hen Egg White Lysozyme
dc.subject
Methylene Carbene
dc.subject
Protein Complexes
dc.subject
Protein Footprinting
dc.subject
Protein Interfaces
dc.subject
Solvent-Accessible Surface Area
dc.subject.classification
Otras Ciencias Biológicas
dc.subject.classification
Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
Exploring protein interfaces with a general photochemical reagent
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2018-03-15T15:23:21Z
dc.identifier.eissn
1469-896X
dc.journal.volume
15
dc.journal.number
4
dc.journal.pagination
744-752
dc.journal.pais
Estados Unidos
dc.journal.ciudad
New York
dc.description.fil
Fil: Gomez, Gabriela Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; Argentina
dc.description.fil
Fil: Cauerhff, Ana. 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
dc.description.fil
Fil: Craig, Patricio Oliver. 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
dc.description.fil
Fil: Goldbaum, Fernando Alberto. 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
dc.description.fil
Fil: Delfino, Jose Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; Argentina
dc.journal.title
Protein Science
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1110/ps.051960406/full
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1110/ps.051960406


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    Articulos de INST.DE QUIMICA Y FISICO-QUIMICA BIOLOGICAS "PROF. ALEJANDRO C. PALADINI"
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    Articulos de INST.DE INVEST.BIOQUIMICAS DE BS.AS(I)

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