INIMEC - CONICET   05467
INSTITUTO DE INVESTIGACION MEDICA MERCEDES Y MARTIN FERREYRA
Unidad Ejecutora - UE
artículos
Título:
Identifying metalloproteins through X-ray fluorescence mapping and mass spectrometry
Autor/es:
RAIMUNDA D.; KHARE T.; GIOMETTI C.; VOGT S.; ARGÜELLO JM.; FINNEY L.
Revista:
METALLOMICS
Editorial:
ROYAL SOC CHEMISTRY
Referencias:
Lugar: CAMBRIDGE; Año: 2012 p. 921 - 927
ISSN:
1756-5901
Resumen:
Metals are critical and dynamic components of biochemistry. To understand their roles, we greatly need tools to identify the ligands that bind them within the complexity of natural systems. This work describes the development of methods that not only detect and distinguish metals, but also characterize the proteins that bind them. We describe an approach to expose, identify and quantify metalloproteins in complex mixtures by sequential non-denaturing 2D-gel electrophoresis/X-ray Fluorescence (XRF) and tandem mass spectrometry (MS/MS) in the same spot of a sample. We first apply the development of 2D GE/XRF to Shewanella oneidensis MR-1, a well-studied system, and verify our electrophoretic approach. Then, we identified a novel periplasmic zinc protein in Pseudomonas aeruginosa PAO1 through 2D-GE/XRF followed by MS/ MS. The identity and function of this protein was verified through a gene mutation experiment.Shewanella oneidensis MR-1, a well-studied system, and verify our electrophoretic approach. Then, we identified a novel periplasmic zinc protein in Pseudomonas aeruginosa PAO1 through 2D-GE/XRF followed by MS/ MS. The identity and function of this protein was verified through a gene mutation experiment.Pseudomonas aeruginosa PAO1 through 2D-GE/XRF followed by MS/ MS. The identity and function of this protein was verified through a gene mutation experiment.