Conformational dynamics of metallo-β-lactamase CcrA during catalysis investigated by using DEER spectroscopy
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| 024 | 7 | 0 | |a 10.1007/s00775-015-1244-8 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00775-015-1244-8 | ||
| 245 | 0 | 0 | |a Conformational dynamics of metallo-β-lactamase CcrA during catalysis investigated by using DEER spectroscopy |h [Elektronische Daten] |c [Mahesh Aitha, Lindsay Moritz, Indra Sahu, Omar Sanyurah, Zahilyn Roche, Robert McCarrick, Gary Lorigan, Brian Bennett, Michael Crowder] |
| 520 | 3 | |a Previous crystallographic and mutagenesis studies have implicated the role of a position-conserved hairpin loop in the metallo-β-lactamases in substrate binding and catalysis. In an effort to probe the motion of that loop during catalysis, rapid-freeze-quench double electron-electron resonance (RFQ-DEER) spectroscopy was used to interrogate metallo-β-lactamase CcrA, which had a spin label at position 49 on the loop and spin labels (at positions 82, 126, or 233) 20-35 Å away from residue 49, during catalysis. At 10ms after mixing, the DEER spectra show distance increases of 7, 10, and 13Å between the spin label at position 49 and the spin labels at positions 82, 126, and 233, respectively. In contrast to previous hypotheses, these data suggest that the loop moves nearly 10Å away from the metal center during catalysis and that the loop does not clamp down on the substrate during catalysis. This study demonstrates that loop motion during catalysis can be interrogated on the millisecond time scale. | |
| 540 | |a SBIC, 2015 | ||
| 690 | 7 | |a Double electron-electron resonance (DEER) |2 nationallicence | |
| 690 | 7 | |a Metallo-β-lactamase (MβLs) |2 nationallicence | |
| 690 | 7 | |a Site-directed spin labeling (SDSL) |2 nationallicence | |
| 690 | 7 | |a Rapid freeze quench (RFQ) |2 nationallicence | |
| 690 | 7 | |a MTSL |2 nationallicence | |
| 690 | 7 | |a CcrA* : Spin-labeledsite-directed variant CcrA:C155S |2 nationallicence | |
| 690 | 7 | |a CcrA*(49) : Spin-labeledsite-directed variant CcrA:C155S/W49C |2 nationallicence | |
| 690 | 7 | |a CcrA*(49/82) : Spin-labeledsite-directed variant CcrA:C155S/W49C/N82C |2 nationallicence | |
| 690 | 7 | |a CcrA*(49/126) : Spin-labeledsite-directed variant CcrA:C155S/W49C/D126C |2 nationallicence | |
| 690 | 7 | |a CcrA*(49/233) : Spin-labeledsite-directed variant CcrA:C155SW49C/E233C |2 nationallicence | |
| 690 | 7 | |a DEER : Double electron-electron resonance/pulsed electron-electron double resonance |2 nationallicence | |
| 690 | 7 | |a EPR : Electron paramagnetic (spin) resonance |2 nationallicence | |
| 690 | 7 | |a MTSL : (S-(1-oxyl-2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrol-3-yl)methyl methanesulfonothioate) |2 nationallicence | |
| 690 | 7 | |a MβL : Metallo-β-lactamase |2 nationallicence | |
| 690 | 7 | |a RFQ : Rapid freeze quench |2 nationallicence | |
| 700 | 1 | |a Aitha |D Mahesh |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | |
| 700 | 1 | |a Moritz |D Lindsay |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | |
| 700 | 1 | |a Sahu |D Indra |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | |
| 700 | 1 | |a Sanyurah |D Omar |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | |
| 700 | 1 | |a Roche |D Zahilyn |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | |
| 700 | 1 | |a McCarrick |D Robert |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | |
| 700 | 1 | |a Lorigan |D Gary |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | |
| 700 | 1 | |a Bennett |D Brian |u Physics Department, Marquette University, 540 N. 15th Street, 53233, Milwaukee, WI, USA |4 aut | |
| 700 | 1 | |a Crowder |D Michael |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | |
| 773 | 0 | |t JBIC Journal of Biological Inorganic Chemistry |d Springer Berlin Heidelberg |g 20/3(2015-04-01), 585-594 |x 0949-8257 |q 20:3<585 |1 2015 |2 20 |o 775 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00775-015-1244-8 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00775-015-1244-8 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Aitha |D Mahesh |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Moritz |D Lindsay |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Sahu |D Indra |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Sanyurah |D Omar |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Roche |D Zahilyn |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a McCarrick |D Robert |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Lorigan |D Gary |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Bennett |D Brian |u Physics Department, Marquette University, 540 N. 15th Street, 53233, Milwaukee, WI, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Crowder |D Michael |u Department of Chemistry and Biochemistry, Miami University, 650 East High Street, 45056, Oxford, OH, USA |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t JBIC Journal of Biological Inorganic Chemistry |d Springer Berlin Heidelberg |g 20/3(2015-04-01), 585-594 |x 0949-8257 |q 20:3<585 |1 2015 |2 20 |o 775 | ||