ICPMS analysis of proteins separated by Native-PAGE: Evaluation of metaloprotein profiles in human synovial fluid with acute and chronic arthritis.
The role of trace elements bound to proteins in the etiology and pathogenesis of rheumatoid arthritis (RA) remains unclear. In this sense, the identification and detection of metalloproteins has a strong and growing interest. Metalloprotein studies are currently carried out by polyacrylamide gel electrophoresis (PAGE) associated to inductively coupled plasma mass spectrometry (ICPMS), and despite that complete information can be obtained for metals such as Fe, Cu and Zn, difficulties due to poor sensitivity for other trace elements such as Sn, As, etc, are currently faced. In the present work, a simple and fast method for the determination of trace metals bound to synovial fluid (SF) proteins was optimized. Proteins from SF (long and short-term RA) were separated in ten fractions by native PAGE, then dissolved in nitric acid and peroxide hydrogen, and analyzed by ICPMS. Fifteen metals were determined in each separated protein fraction (band). Adequate calibration of proteins molecular weight allowed stablishing which protein type were bound to different metals.
Instituto de Química San Luis (INQUISAL-CONICET), Área de Química Analítica, Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis. San Luis 5700 Argentina.
Mario F
MF
Moyano
Instituto de Química San Luis (INQUISAL-CONICET), Área de Química Analítica, Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis. San Luis 5700 Argentina.
Leonardo
L
Mariño-Repizo
Clínica Bolivar, Bolivar 1277, Ciudad de San Luis, 5700 Argentina, Argentina.
Héctor
H
Tamashiro
Instituto de Química San Luis (INQUISAL-CONICET), Área de Química Analítica, Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis. San Luis 5700 Argentina.
Liliana
L
Villegas
Instituto de Química San Luis (INQUISAL-CONICET), Área de Química Analítica, Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis. San Luis 5700 Argentina.
Mariano
M
Acosta
Instituto de Química San Luis (INQUISAL-CONICET), Área de Química Analítica, Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis. San Luis 5700 Argentina. Electronic address: ragil@unsl.edu.ar.
Raúl A
RA
Gil
D000208:Acute Disease; D001172:Arthritis, Rheumatoid / Q000378:metabolism*; D002908:Chronic Disease; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D013058:Mass Spectrometry; D008667:Metalloproteins / Q000032:analysis*; D013582:Synovial Fluid / Q000737:chemistry*
D008667:Metalloproteins
Human synovial fluid proteins; Inductively coupled plasma mass spectrometry; Polyacrlylamide gel electrophoresis; Trace elements