Artículo
The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins
Ching, H. Y. Vincent; Mascali, Florencia Carla
; Bertrand, Hélène C.; Bruch, Eduardo Marcos
; Demay Drouhard, Paul; Rasia, Rodolfo Maximiliano
; Policar, Clotilde; Tabares, Leandro Cesar
; Un, Sun
Fecha de publicación:
03/2016
Editorial:
American Chemical Society
Revista:
Journal of Physical Chemistry Letters
ISSN:
1948-7185
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
A genetically encodable paramagnetic spin-label capable of self-assembly from naturally available components would offer a means for studying the in-cell structure and interactions of a protein by electron paramagnetic resonance (EPR). Here, we demonstrate pulse electron-electron double resonance (DEER) measurements on spin-labels consisting of Mn(II) ions coordinated to a sequence of histidines, so-called His-tags, that are ubiquitously added by genetic engineering to facilitate protein purification. Although the affinity of His-tags for Mn(II) was low (800 μM), Mn(II)-bound His-tags yielded readily detectable DEER time traces even at concentrations expected in cells. We were able to determine accurately the distance between two His-tag Mn(II) spin-labels at the ends of a rigid helical polyproline peptide of known structure, as well as at the ends of a completely cell-synthesized 3-helix bundle. This approach not only greatly simplifies the labeling procedure but also represents a first step towards using self-assembling metal spin-labels for in-cell distance measurements.
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Articulos(IBR)
Articulos de INST.DE BIOLOGIA MOLECULAR Y CELULAR DE ROSARIO
Articulos de INST.DE BIOLOGIA MOLECULAR Y CELULAR DE ROSARIO
Citación
Ching, H. Y. Vincent; Mascali, Florencia Carla; Bertrand, Hélène C.; Bruch, Eduardo Marcos; Demay Drouhard, Paul; et al.; The Use of Mn(II) Bound to His-tags as Genetically Encodable Spin-Label for Nanometric Distance Determination in Proteins; American Chemical Society; Journal of Physical Chemistry Letters; 7; 6; 3-2016; 1072-1076
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