Identification of a metallothionein in Synechococcus by capillary electrophoresis hyphenated with inductively coupled plasma mass spectrometry.
A home-made system hyphenating capillary electrophoresis with an inductively coupled plasma mass spectrometer (CE-ICP-MS) for cadmium speciation of protein-binding and free cadmium ions in solution is presented. The CE-ICP-MS interface consisted of an acrylic block with an internal volume ca. 20 microL in which a platinum electrode, a capillary column, and a connection to an ICP nebulizer were inserted. A make-up electrolyte solution containing 50 mmol L(-1) Tris-HCl buffer solution (pH 9.0) was continuously flowed through the interface to the ICP nebulizer. The separation of free Cd ions, Cd-cysteine, and Cd bounded to metallothionein (MT) isoforms from rabbit liver was carried out by capillary electrophoresis, and the analytes were detected by ICP-MS. The feasibility to isolate metallothionein compounds extracted from the cyanobacterium Synechococcus PCC7942 was demonstrated. The Cd binding proteins were induced in Synechococcus PCC7942 and further analyzed by CE ICP-MS.
Lavorante
André F
AF
Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, Av. Centenário 303, 13416-000, Piracicaba, SP, Brazil.
Giné
Maria Fernanda
MF
Gervasio
Ana Paula G
AP
Miranda
Carlos E S
CE
Fiore
Marli F
MF
Bellato
Cláudia M
CM
Carrilho
Emanuel
E
D000818:Animals; D002104:Cadmium / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D000458:Cyanobacteria / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D008099:Liver / Q000737:chemistry; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D008668:Metallothionein / Q000302:isolation & purification* / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits
D002104:Cadmium; D008668:Metallothionein