ICPMS analysis of proteins separated by Native-PAGE: Evaluation of metaloprotein profiles in human synovial fluid with acute and chronic arthritis

MARIO F. MOYANO; LEONARDO MARIÑO-REPIZO; HÉCTOR TAMASHIRO; LILIANA VILLEGAS; MARIANO ACOSTA; RAÚL A. GIL

JOURNAL OF TRACE ELEMENTS IN MEDICINE AND BIOLOGY

ELSEVIER GMBH

Año: 2016 vol. 36 p. 44 - 51

0946-672X

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 andgrowing interest. Metalloprotein studies are currently carried out by polyacrylamide gel electrophoresis(PAGE) associated to inductively coupled plasma mass spectrometry (ICPMS), and despite that completeinformation can be obtained for metals such as Fe, Cu and Zn, difficulties due to poor sensitivity for othertrace elements such as Sn, As, etc, are currently faced. In the present work, a simple and fast method forthe 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 andperoxide hydrogen, and analyzed by ICPMS. Fifteen metals were determined in each separated proteinfraction (band). Adequate calibration of proteins molecular weight allowed stablishing which proteintype were bound to different metals.