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Although extensively\ncharacterized from the spectroscopic and structural points\u2010of\u2010view, some of its kinetic aspects\nare still under explored. In this work the kinetic behaviour of ccNiR has been evaluated in a\nsystematic manner using two different spectrophotometric assays carried out in the presence of\ndifferent redox mediators and a direct electrochemical approach. Solution assays have proved\nthat the specific activity of ccNiR decreases with the reduction potential of the electronic carriers\nand ammonium is always the main product of nitrite reduction. The catalytic parameters were\ndiscussed on the basis of the mediator reducing power and also taking into account the location\nof their putative docking sites with ccNiR. Due to the fast kinetics of ccNiR, electron delivering\nfrom reduced electron donors is rate\u2010limiting in all spectrophotometric assays, so the estimated\nkinetic constants are apparent only. Nevertheless, this limitation could be overcome by using a\ndirect electrochemical approach which shows that the binding affinity for nitrite decreases whilst\nturnover increases with the reductive driving force.<\/jats:p>","DOI":"10.1155\/2010\/634597","type":"journal-article","created":{"date-parts":[[2010,6,22]],"date-time":"2010-06-22T14:35:29Z","timestamp":1277217329000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Measuring the Cytochrome <i>c<\/i> Nitrite Reductase Activity\u2014Practical Considerations on the Enzyme Assays"],"prefix":"10.1155","volume":"2010","author":[{"given":"C\u00e9lia M.","family":"Silveira","sequence":"first","affiliation":[]},{"given":"St\u00e9phane","family":"Besson","sequence":"additional","affiliation":[]},{"given":"Isabel","family":"Moura","sequence":"additional","affiliation":[]},{"given":"Jos\u00e9 J. 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NussinovR. andWolfsonH. GuigoR.andGusfieldD. Efficient unbound docking of rigid molecules 2452 Proceedings of the 2nd Workshop on Algorithms in Bioinformatics(WABI \u203202) 2002 Rome Italy Springer Lecture Notes in Computer Science.","DOI":"10.1007\/3-540-45784-4_14"},{"key":"e_1_2_6_28_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gki481"},{"key":"e_1_2_6_29_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.20084"},{"key":"e_1_2_6_30_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2006.12.023"},{"key":"e_1_2_6_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167\u20104838(00)00111\u20104"},{"key":"e_1_2_6_32_2","volume-title":"Enzyme Assays: A Practical Approach","author":"Tipton K. F.","year":"1993"},{"key":"e_1_2_6_33_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja054160s"},{"key":"e_1_2_6_34_2","volume-title":"Fundamentals of Electroanalytical Chemistry","author":"Monk P. M. 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