Cleaning protocols for crystallization robots: Preventing protease contamination

Andreas Naschberger; Barbara G. Fürnrohr; Theresia Dunzendorfer-Matt; Christopher A. Bonagura; David Wright; Klaus Scheffzek; Bernhard Rupp

doi:10.1107/S2053230X14026053

Cleaning protocols for crystallization robots: Preventing protease contamination

Andreas Naschberger, Barbara G. Fürnrohr, Theresia Dunzendorfer-Matt, Christopher A. Bonagura, David Wright, Klaus Scheffzek, Bernhard Rupp

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

The protease in the commonly used commercial low-foam enzyme cleaner Zymit cannot be completely blocked by EDTA, a widely used inhibitor of metalloproteases, at concentrations of up to 5mM. Severe protein degradation was observed in crystallization drops after EDTA-containing wash steps unless residual Zymit protease was removed with NaOH at a concentration of at least 0.1M. Wash steps with 0.1% SDS were also ineffective in completely removing the remaining Zymit activity. Protocols including wash steps with at least 0.1M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity.

Original language	English (US)
Pages (from-to)	100-102
Number of pages	3
Journal	Acta Crystallographica Section F:Structural Biology Communications
Volume	71
DOIs	https://doi.org/10.1107/S2053230X14026053
State	Published - Jan 1 2015
Externally published	Yes

Bibliographical note

Generated from Scopus record by KAUST IRTS on 2023-02-15

ASJC Scopus subject areas

Biophysics
Condensed Matter Physics
Genetics
Biochemistry
Structural Biology
Medicine(all)

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Cite this

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title = "Cleaning protocols for crystallization robots: Preventing protease contamination",

abstract = "The protease in the commonly used commercial low-foam enzyme cleaner Zymit cannot be completely blocked by EDTA, a widely used inhibitor of metalloproteases, at concentrations of up to 5mM. Severe protein degradation was observed in crystallization drops after EDTA-containing wash steps unless residual Zymit protease was removed with NaOH at a concentration of at least 0.1M. Wash steps with 0.1% SDS were also ineffective in completely removing the remaining Zymit activity. Protocols including wash steps with at least 0.1M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity.",

author = "Andreas Naschberger and F{\"u}rnrohr, {Barbara G.} and Theresia Dunzendorfer-Matt and Bonagura, {Christopher A.} and David Wright and Klaus Scheffzek and Bernhard Rupp",

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doi = "10.1107/S2053230X14026053",

language = "English (US)",

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T1 - Cleaning protocols for crystallization robots: Preventing protease contamination

AU - Naschberger, Andreas

AU - Fürnrohr, Barbara G.

AU - Dunzendorfer-Matt, Theresia

AU - Bonagura, Christopher A.

AU - Wright, David

AU - Scheffzek, Klaus

AU - Rupp, Bernhard

N1 - Generated from Scopus record by KAUST IRTS on 2023-02-15

PY - 2015/1/1

Y1 - 2015/1/1

N2 - The protease in the commonly used commercial low-foam enzyme cleaner Zymit cannot be completely blocked by EDTA, a widely used inhibitor of metalloproteases, at concentrations of up to 5mM. Severe protein degradation was observed in crystallization drops after EDTA-containing wash steps unless residual Zymit protease was removed with NaOH at a concentration of at least 0.1M. Wash steps with 0.1% SDS were also ineffective in completely removing the remaining Zymit activity. Protocols including wash steps with at least 0.1M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity.

AB - The protease in the commonly used commercial low-foam enzyme cleaner Zymit cannot be completely blocked by EDTA, a widely used inhibitor of metalloproteases, at concentrations of up to 5mM. Severe protein degradation was observed in crystallization drops after EDTA-containing wash steps unless residual Zymit protease was removed with NaOH at a concentration of at least 0.1M. Wash steps with 0.1% SDS were also ineffective in completely removing the remaining Zymit activity. Protocols including wash steps with at least 0.1M NaOH, as for example specified in the original ZENM protocol, are recommended to completely deactivate Zymit protease activity.

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DO - 10.1107/S2053230X14026053

M3 - Article

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JO - Acta Crystallographica Section F:Structural Biology Communications

JF - Acta Crystallographica Section F:Structural Biology Communications

ER -

Cleaning protocols for crystallization robots: Preventing protease contamination

Abstract

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