Assessment of metals bound to marine plankton proteins and to dissolved proteins in seawater

• Published in: Analytica chimica acta Vol. 804; pp. 59 - 65
• Main Authors: García-Otero, Natalia, Barciela-Alonso, María Carmen, Bermejo-Barrera, Pilar, Moreda-Piñeiro, Antonio, Jiménez, María S., Gómez, María T., Castillo, Juan R.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 04.12.2013
• Subjects: 2D polyacrylamide gel electrophoresis; Cadmium; Chromium; Copper; Dissolution; Dissolved proteins; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Laser ablation inductively coupled plasma mass spectrometry; Marine; Marine Biology; Mass Spectrometry; Metal-binding proteins; Metals - metabolism; Methyl alcohol; Molecular weight; Phytoplankton; Plankton; Plankton - metabolism; Plant Proteins - metabolism; Proteins; Seawater; Ultrafiltration; Dissolved proteins; Metal-binding proteins; Phytoplankton; 2D polyacrylamide gel electrophoresis; Laser ablation inductively coupled plasma mass spectrometry
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2013.10.016

•Study of the metals bound to marine proteins in seawater and marine phytoplankton.•Use of the 2D PAGE and LA–ICP-MS in the study of marine proteins.•The presence of metals in dissolved marine proteins was demonstrated by the first time. Studies based on laser ablation–inductively coupled plasma-mass spectrometry (LA–ICP-MS) have been performed to assess metal bound to dissolved proteins and proteins from marine plankton after two-dimensional polyacrylamide gel electrophoresis (2D PAGE). Dissolved proteins were pre-concentrated from surface seawater (60L) by tangential ultrafiltration with 10kDa molecular weight cut-off (MWCO) membranes and further centrifugal ultrafiltration (10kDa) before proteins isolation by methanol/chloroform/water precipitation. Proteins isolation from plankton was assessed after different trichloroacetic acid (TCA)/acetone and methanol washing stages, and further proteins extraction with a phenol solution. LA–ICP-MS analysis of the electrophoretic profiles obtained for dissolved proteins shows the presence of Cd, Cr, Cu, and Zn in five spots analyzed. These proteins exhibit quite similar molecular weights (within the 10–14kDa range) and pIs (from 5.8 to 7.3). Cd, Cr, Cu, and Zn have also been found to be associated to proteins isolated from plankton samples. In this case, Cd has been found to be bound to proteins of quite different molecular weight (9, 13 and 22kDa) and pIs (4.5, 5.2, 5.5, and 10). However, trace elements such as Cr, Cu and Zn appear to be mainly bound to plankton proteins of low molecular weight and variable pI.