Mechanism of the binding of Z-L-tryptophan and Z-L-phenylalanine to thermolysin and stromelysin-1 in aqueous solutions

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Show simple item record Ceruso, Mariangela Howe, Nicole Malthouse, J.Paul G. 2012-01-30T12:51:57Z 2012-01-30T12:51:57Z 2011 Elsevier B.V. en 2012-02
dc.identifier.citation Biochimica et Biophysica Acta (BBA) - Proteins & Proteomics en
dc.identifier.issn 1570-9639
dc.description.abstract The chemical shift of the carboxylate carbon of Z-tryptophan is increased from 179.85 to 182.82 ppm and 182.87 on binding to thermolysin and stromelysin-1 respectively. The chemical shift of Z-phenylalanine is also increased from 179.5 ppm to 182.9 ppm on binding to thermolysin. From pH studies we conclude that the pKa of the inhibitor carboxylate group is lowered by at least 1.5 pKa units when it binds to either enzyme. The signal at ~183 ppm is no longer observed when the active site zinc atom of thermolysin or stromelysin-1 is replaced by cobalt. We estimate that the distance of carboxylate carbon of Z-[1-13C]-L-tryptophan is ≤ 3.71 Å from the active site cobalt atom of thermolysin. We conclude that the side chain of Z-[1-13C]-L-tryptophan is not bound in the S2' subsite of thermolysin. As the chemical shifts of the carboxylate carbons of the bound inhibitors are all ~183 ppm we conclude that they are all bound in a similar way most probably with the inhibitor carboxylate group directly coordinated to the active site zinc atom. Our spectrophotometric results confirm that the active site zinc atom is tetrahedrally coordinated when the inhibitors Z-tryptophan or Z-phenylalanine are bound to thermolysin. en
dc.description.sponsorship Science Foundation Ireland en
dc.description.sponsorship Higher Education Authority en
dc.description.sponsorship Other funder en
dc.format.extent 2101793 bytes
dc.format.mimetype application/pdf
dc.language.iso en en
dc.publisher Elsevier B.V. en
dc.rights This is the author’s version of a work that was accepted for publication in Biochimica et Biophysica Acta (BBA) - Proteins & Proteomics. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Biochimica et Biophysica Acta (BBA) - Proteins & Proteomics 1824 : 303-310 DOI: 10.1016/j.bbapap.2011.10.007 en
dc.subject Thermolysin en
dc.subject Stromelysin en
dc.subject NMR en
dc.subject Amino acids en
dc.subject Binding en
dc.subject Cobalt en
dc.subject.lcsh Metalloproteinases en
dc.subject.lcsh Amino acids en
dc.subject.lcsh Nuclear magnetic resonance en
dc.subject.lcsh Cobalt en
dc.title Mechanism of the binding of Z-L-tryptophan and Z-L-phenylalanine to thermolysin and stromelysin-1 in aqueous solutions en
dc.type Journal Article en
dc.internal.availability Full text available en
dc.internal.webversions en
dc.status Peer reviewed en
dc.identifier.volume 1824 en
dc.identifier.issue 2 en
dc.identifier.startpage 303 en
dc.identifier.endpage 310 en
dc.identifier.doi 10.1016/j.bbapap.2011.10.007
dc.neeo.contributor Ceruso|Mariangela|aut| en
dc.neeo.contributor Howe|Nicole|aut| en
dc.neeo.contributor Malthouse|J.Paul G.|aut| en
dc.description.othersponsorship Wellcome Trust en
dc.description.othersponsorship University College Dublin. President’s Research Fellowship en
dc.description.admin ke, sp, - kpw22/12/11 en

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