caa a22 4500 477136427 CHVBK 20180405111719.0 cr unu---uuuuu 170330e19970401xx s 000 0 eng 10.1023/A:1018683026421 doi (NATIONALLICENCE)springer-10.1023/A:1018683026421 Selective observation of the Cu(I)-amicyanin metal site by paramagnetic NMR on partially oxidised samples [Elektronische Daten] [Jesús Salgado, Arnout Kalverda, Gerard Canters] The relaxation enhancement caused by paramagnetic copper(II) is used to observe selectivelythe metal site of copper(I)-amicyanin by one- and two-dimensional NMR spectroscopy. Theparamagnetic effect is communicated to the diamagnetic protein through the electron self-exchange reaction in partially oxidised samples, and can be used for the selective detectionof protons around the metal. Relaxation-selective NMR pulse sequences, like super-WEFTand WEFT-NOESY, are used to achieve the desired selection of the signals. The spectraobtained show well-resolved signals corresponding to protons within a radius of∼7 Å from the metal, including almost all protons from the coordinated residues. A significant increasein resolution as well as selection of the most relevant part of the protein (close to the activecentre) are the principal advantages of this technique, which can be used to obtain specificinformation about the metal site in blue copper proteins, to assist in the assignment of theirNMR spectra and to determine functional properties like the electron self-exchange rate. Kluwer Academic Publishers, 1997 Copper proteins nationallicence Type 1 copper site nationallicence Azurin nationallicence Electron transfer nationallicence Paramagnetic relaxation nationallicence Spectroscopic probe nationallicence Salgado Jesús Leiden Institute of Chemistry, Gorlaeus Laboratories, University of Leiden, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut Kalverda Arnout Leiden Institute of Chemistry, Gorlaeus Laboratories, University of Leiden, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut Canters Gerard Leiden Institute of Chemistry, Gorlaeus Laboratories, University of Leiden, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut Journal of Biomolecular NMR Kluwer Academic Publishers 9/3(1997-04-01), 299-305 0925-2738 9:3<299 1997 9 10858 https://doi.org/10.1023/A:1018683026421 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1018683026421 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Salgado Jesús Leiden Institute of Chemistry, Gorlaeus Laboratories, University of Leiden, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut NATIONALLICENCE 700 1- Kalverda Arnout Leiden Institute of Chemistry, Gorlaeus Laboratories, University of Leiden, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut NATIONALLICENCE 700 1- Canters Gerard Leiden Institute of Chemistry, Gorlaeus Laboratories, University of Leiden, P.O. Box 9502, 2300 RA, Leiden, The Netherlands aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Kluwer Academic Publishers 9/3(1997-04-01), 299-305 0925-2738 9:3<299 1997 9 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer