10.1002\/0470862106.ia340<\/a><\/p>\nAuthors:<\/h4>\n
Simon de Vries<\/p>\n
Abstract:<\/h4>\n
Abstract Rapid\u2010mixing, rapid\u2010freezing techniques are employed for the study of enzyme or chemical catalytic mechanisms. The techniques aim to provide electronic and molecular structures of transient intermediates formed during the reaction by means of spectroscopic analyses of the rapidly frozen samples. Briefly, the mixing\u2010sampling procedure consists of the rapid mixing of two reactants, for example, enzyme and substrate, followed by rapid freezing of the mixture in a liquid cryo\u2010medium or on a cold plate to quench the reaction progress. The resulting frozen powder is subsequently sampled and made available for further low\u2010temperature spectroscopic analyses such as UV\u2010Vis spectroscopy, multifrequency Electron Paramagnetic Resonance (EPR) spectroscopy, Electron Nuclear Double Resonance (ENDOR) spectroscopy, Electron Spin Echo Envelope Modulation (ESEEM), Magnetic Circular Dichro\u00efsm (MCD) spectroscopy, M\u00f6ssbauer spectroscopy, resonance Raman spectroscopy, or X\u2010ray Absorption Spectroscopy. By preparing a series of samples freeze\u2010quenched after various times, a full kinetic profile of the catalytic cycle of an enzyme or any other chemical reaction is obtained. The structures of intermediates are assigned in conjunction with (a selection of) the spectroscopic techniques listed above. This article details various aspects of continuous\u2010flow and stopped\u2010flow rapid\u2010mixing techniques, considering mixer design, freeze\u2010quenching methodology, and sampling. The scope and limitations to (bio)chemical research of the well\u2010established rapid freeze\u2010quench (RFQ) and the recently developed microsecond freeze\u2010hyperquenching (MHQ) technologies are specifically highlighted.<\/p>\n","protected":false},"excerpt":{"rendered":"
DOI: 10.1002\/0470862106.ia340 Authors: Simon de Vries Abstract: Abstract Rapid\u2010mixing, rapid\u2010freezing techniques are employed for the study of enzyme or chemical catalytic mechanisms. The techniques aim to provide electronic and molecular structures of transient intermediates formed during the reaction by means of spectroscopic analyses of the rapidly frozen samples. Briefly, the mixing\u2010sampling procedure consists of the […]<\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[5],"tags":[],"class_list":["post-645","post","type-post","status-publish","format-standard","hentry","category-freeze-quench"],"_links":{"self":[{"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=\/wp\/v2\/posts\/645","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=645"}],"version-history":[{"count":0,"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=\/wp\/v2\/posts\/645\/revisions"}],"wp:attachment":[{"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=645"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=645"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/citations.tools.bio-logic.fr\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=645"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}