MAD phasing with krypton
Experiments demonstrating the feasibility of Kr-edge MAD on frozen crystals as a routine method for structure determination are reported. Approximately 50 % of protein crystals can be successfully derivatized by pressurization with the noble gases xenon or krypton. While Xe has produced many useful...
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ftciteseerx:oai:CiteSeerX.psu:10.1.1.507.8909 2023-05-15T18:26:46+02:00 MAD phasing with krypton Aina Cohen A Paul Ellis A Nicole S. Michael Soltisa The Pennsylvania State University CiteSeerX Archives application/pdf http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.507.8909 http://smb.slac.stanford.edu/research/staff_publications/kr_reprint.pdf en eng http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.507.8909 http://smb.slac.stanford.edu/research/staff_publications/kr_reprint.pdf Metadata may be used without restrictions as long as the oai identifier remains attached to it. http://smb.slac.stanford.edu/research/staff_publications/kr_reprint.pdf text ftciteseerx 2016-01-08T09:28:05Z Experiments demonstrating the feasibility of Kr-edge MAD on frozen crystals as a routine method for structure determination are reported. Approximately 50 % of protein crystals can be successfully derivatized by pressurization with the noble gases xenon or krypton. While Xe has produced many useful derivatives for MIR phasing over the last several years, the Xe edges (K edge = 34.6 keV, LI = 5.5 keV) are not easily accessible for MAD studies. As the Kr K edge (14.3 keV) is accessible on most MAD beamlines, Kr derivatization provides the additional opportunity to conduct a MAD experiment and obtain phases using only a single crystal. This paper describes the phasing of two proteins using Kr MAD: the 17 kDa Fe protein myoglobin (Mb) from sperm whale (Physeter catodon) and an 18 kDa protein (SP18) from green abalone (Haliotis fulgens). Three-wavelength data were collected at SSRL beamline 9-2 from crystals of Mb and SP18 incubated in 2.76 MPa of Kr gas for 2 min, depressurized and then ¯ash-frozen in a stream of nitrogen gas at 100 K. MAD phases were calculated using the program SHARP and the resulting density improved with wARP. The ®nal maps for both Mb and SP18 were of excellent quality. Text Sperm whale Unknown Catodon ENVELOPE(-59.966,-59.966,-63.500,-63.500) |
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ftciteseerx |
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English |
description |
Experiments demonstrating the feasibility of Kr-edge MAD on frozen crystals as a routine method for structure determination are reported. Approximately 50 % of protein crystals can be successfully derivatized by pressurization with the noble gases xenon or krypton. While Xe has produced many useful derivatives for MIR phasing over the last several years, the Xe edges (K edge = 34.6 keV, LI = 5.5 keV) are not easily accessible for MAD studies. As the Kr K edge (14.3 keV) is accessible on most MAD beamlines, Kr derivatization provides the additional opportunity to conduct a MAD experiment and obtain phases using only a single crystal. This paper describes the phasing of two proteins using Kr MAD: the 17 kDa Fe protein myoglobin (Mb) from sperm whale (Physeter catodon) and an 18 kDa protein (SP18) from green abalone (Haliotis fulgens). Three-wavelength data were collected at SSRL beamline 9-2 from crystals of Mb and SP18 incubated in 2.76 MPa of Kr gas for 2 min, depressurized and then ¯ash-frozen in a stream of nitrogen gas at 100 K. MAD phases were calculated using the program SHARP and the resulting density improved with wARP. The ®nal maps for both Mb and SP18 were of excellent quality. |
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The Pennsylvania State University CiteSeerX Archives |
format |
Text |
author |
Aina Cohen A Paul Ellis A Nicole S. Michael Soltisa |
spellingShingle |
Aina Cohen A Paul Ellis A Nicole S. Michael Soltisa MAD phasing with krypton |
author_facet |
Aina Cohen A Paul Ellis A Nicole S. Michael Soltisa |
author_sort |
Aina Cohen |
title |
MAD phasing with krypton |
title_short |
MAD phasing with krypton |
title_full |
MAD phasing with krypton |
title_fullStr |
MAD phasing with krypton |
title_full_unstemmed |
MAD phasing with krypton |
title_sort |
mad phasing with krypton |
url |
http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.507.8909 http://smb.slac.stanford.edu/research/staff_publications/kr_reprint.pdf |
long_lat |
ENVELOPE(-59.966,-59.966,-63.500,-63.500) |
geographic |
Catodon |
geographic_facet |
Catodon |
genre |
Sperm whale |
genre_facet |
Sperm whale |
op_source |
http://smb.slac.stanford.edu/research/staff_publications/kr_reprint.pdf |
op_relation |
http://citeseerx.ist.psu.edu/viewdoc/summary?doi=10.1.1.507.8909 http://smb.slac.stanford.edu/research/staff_publications/kr_reprint.pdf |
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Metadata may be used without restrictions as long as the oai identifier remains attached to it. |
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1766208739599187968 |