دورية أكاديمية
Exploring protein interfaces with a general photochemical reagent
| العنوان: | Exploring protein interfaces with a general photochemical reagent |
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| المؤلفون: | Gomez, Gabriela Elena, Cauerhff, Ana, Craig, Patricio Oliver, Goldbaum, Fernando Alberto, Delfino, Jose Maria |
| بيانات النشر: | Cold Spring Harbor Lab Press |
| المجموعة: | CONICET Digital (Consejo Nacional de Investigaciones Científicas y Técnicas) |
| مصطلحات موضوعية: | Diazirine, Epitope Mapping, Hen Egg White Lysozyme, Methylene Carbene, Protein Complexes, Protein Footprinting, Protein Interfaces, Solvent-Accessible Surface Area, https://purl.org/becyt/ford/1.6Test, https://purl.org/becyt/ford/1Test |
| الوصف: | 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. ; Fil: Gomez, Gabriela Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas ; Argentina ; Fil: Cauerhff, Ana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación ... |
| نوع الوثيقة: | article in journal/newspaper |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 0961-8368 1469-896X |
| العلاقة: | http://hdl.handle.net/11336/39137Test; 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; CONICET Digital; CONICET |
| الإتاحة: | https://doi.org/10.1110/ps.051960406Test http://hdl.handle.net/11336/39137Test |
| حقوق: | info:eu-repo/semantics/openAccess ; https://creativecommons.org/licenses/by-nc-sa/2.5/arTest/ |
| رقم الانضمام: | edsbas.45CF6B21 |
| قاعدة البيانات: | BASE |
| تدمد: | 09618368 1469896X |
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