Separative techniques for metalloproteomics require balance between separation and perturbation

Development of a metalloproteomic approach to analyse the response of Arabidopsis cells to uranium stress

“…protein tandem mass spectrometry (MS/MS) and inductively coupled plasma mass spectrometry (ICP-MS), respectively. 24,25 Metalloproteins account for more than a third of total proteins and are involved in a wide array of functions related to the functional, structural and regulatory roles of bound metals. 26,27 The main challenge in the identification of metalloproteins by using topdown proteomic approaches is to preserve non-covalent and labile metal-protein complexes during protein fractionation steps.…”

Cadmium specific proteomic responses of a highly resistantPseudomonas aeruginosasan ai

“…8,9,11,12 In order to enrich metalloproteins in an analyzed proteome, size exclusion chromatography (SEC) has been used for protein prefractionation. 15 SEC has been reliable in separation of metalloproteins and protein complexes maintaining them in a native state. 15,16 P. aeruginosa san ai, an environmental isolate from mineral cutting oil, was previously used in various studies in our laboratory related to rhamnolipid, 17 exopolysaccharide (EPS), 18 the extracellular enzymes, lipase and protease, 19,20 and their application in detergents 21 as well as in the study related to the organism's capacity for chromium removal.…”

“…15 SEC has been reliable in separation of metalloproteins and protein complexes maintaining them in a native state. 15,16 P. aeruginosa san ai, an environmental isolate from mineral cutting oil, was previously used in various studies in our laboratory related to rhamnolipid, 17 exopolysaccharide (EPS), 18 the extracellular enzymes, lipase and protease, 19,20 and their application in detergents 21 as well as in the study related to the organism's capacity for chromium removal. 22 In this study, we investigated the distribution of cadmium, its uptake by extracellular polymers and effects of intracellular cadmium on protein expression in P. aeruginosa san ai.…”

Assessment of CE‐ICP/MS hyphenation for the study of uranyl/protein interactions

“…Second, the separation conditions dictated by the need of a high resolution often lead to the disruption of metal–protein interactions. Several reviews discussed these advantages and drawbacks . Although SEC and CE have to cope with a low resolution, these techniques appear to be good options, less intrusive than others, since no chemical interactions theoretically occur during the separation.…”