000129813 001__ 129813
000129813 005__ 20240111114817.0
000129813 0247_ $$2doi$$a10.1007/s00216-012-6461-1
000129813 0248_ $$2sideral$$a78770
000129813 037__ $$aART-2013-78770
000129813 041__ $$aeng
000129813 100__ $$0(orcid)0000-0002-8460-6020$$aJiménez, M.S.$$uUniversidad de Zaragoza
000129813 245__ $$aEvaluation of gel electrophoresis techniques and laser ablation–inductively coupled plasma-mass spectrometry for screening analysis of Zn and Cu-binding proteins in plankton
000129813 260__ $$c2013
000129813 5060_ $$aAccess copy available to the general public$$fUnrestricted
000129813 5203_ $$aThe determination of metal-binding proteins in plankton is important because of their involvement in photosynthesis, which is fundamental to the biogeochemical cycle of the oceans and other ecosystems. We have elaborated a new strategy for screening of Cu and Zn-containing proteins in plankton on the basis of separation of proteins by use of Blue-Native PAGE (BN-PAGE), which entails use of a non-denaturing Tris–tricine system and detection of metals in the proteins by laser ablation inductively coupled plasma mass spectrometry (LA–ICP–MS). For comparison, denaturing PAGE based on Tris–glycine and Tris–tricine systems and Anodic-Native PAGE have also been investigated. A large number of protein bands with MW between 20 and 75 kDa were obtained by use of Tris–glycine PAGE but detection of metals by LA–ICP–MS was unsuccessful because of loss of metals from the proteins during the separation process. Different protein extraction, purification, and preconcentration methods were evaluated, focussing on both issues—achieving the best extraction and characterization of the proteins while maintaining the integrity of metal–protein binding in the plankton sample. Use of 25 mmol L−1 Tris–HCl and a protease inhibitor as extraction buffer with subsequent ultrafiltration and acetone precipitation was the most efficient means of sample preparation. Two Cu and Zn proteins were detected, a protein band corresponding to a MW of 60 kDa and another poorly resolved band with a MW between 15 and 35 kDa.
000129813 540__ $$9info:eu-repo/semantics/openAccess$$aAll rights reserved$$uhttp://www.europeana.eu/rights/rr-f/
000129813 590__ $$a3.578$$b2013
000129813 591__ $$aCHEMISTRY, ANALYTICAL$$b11 / 75 = 0.147$$c2013$$dQ1$$eT1
000129813 591__ $$aBIOCHEMICAL RESEARCH METHODS$$b21 / 77 = 0.273$$c2013$$dQ2$$eT1
000129813 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/acceptedVersion
000129813 700__ $$aRodriguez, L.
000129813 700__ $$0(orcid)0000-0001-6673-489X$$aBertolin, J.R.
000129813 700__ $$0(orcid)0000-0003-1400-9041$$aGómez, M.T.$$uUniversidad de Zaragoza
000129813 700__ $$0(orcid)0000-0002-4070-8607$$aCastillo, J.R.$$uUniversidad de Zaragoza
000129813 7102_ $$12009$$2750$$aUniversidad de Zaragoza$$bDpto. Química Analítica$$cÁrea Química Analítica
000129813 773__ $$g405, 1 (2013), 359-368$$pAnal. Bioanal. Chem.$$tAnalytical and Bioanalytical Chemistry$$x1618-2642
000129813 8564_ $$s713494$$uhttps://zaguan.unizar.es/record/129813/files/texto_completo.pdf$$yPostprint
000129813 8564_ $$s808886$$uhttps://zaguan.unizar.es/record/129813/files/texto_completo.jpg?subformat=icon$$xicon$$yPostprint
000129813 909CO $$ooai:zaguan.unizar.es:129813$$particulos$$pdriver
000129813 951__ $$a2024-01-11-09:29:37
000129813 980__ $$aARTICLE