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J Biochem Biophys Methods ; 62(2): 97-109, 2005 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-15680280

RESUMO

In the present work, protein bands from in vitro embriogenic callus (Citrus sinensis L. Osbeck) were investigated using micro-synchrotron radiation X-ray fluorescence (muSR-XRF) after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) separation. Metal-binding protein quantification was done after microwave oven decomposition of gel by synchrotron radiation total reflection X-ray fluorescence (SR-TXRF), flame atomic absorption spectrometry (FAAS) and flame atomic emission spectrometry (FAES). According to the analysis of the protein bands, it is possible to observe that both 81 and ca. 14 kDa proteins present different Fe signal intensity at different positions. The analysis of 53 kDa protein, showed even more interesting results. Besides Fe, the muSR-XRF experiments indicate the presence of Ca, Cu, K and Zn. Chemical elements such as Cu, K, Fe and Zn were determined by SR-TXRF, Mg by FAAS and Na by FAES. Ca was determined by SR-TXRF and FAAS only for accuracy check. In the mineralised protein bands of 81 and around 14 kDa band, only Fe was determined (105 and 21.8 microg g(-1)). For those protein bands (86-ca. 14 kDa) were determined, Ca, K, Cu and Zn in a wide concentration range (42.4-283, 2.47-96.8, 0.91-15.9 and 3.39-29.7 microg g(-1), respectively).


Assuntos
Biofísica/métodos , Metais/química , Espectrofotometria Atômica/métodos , Síncrotrons , Cálcio/análise , Citrus/metabolismo , Cobre/análise , Eletroforese em Gel de Poliacrilamida , Peso Molecular , Potássio/análise , Ligação Proteica , Proteínas/química , Sódio/análise , Espectrometria de Fluorescência , Temperatura , Raios X , Zinco/análise
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