Yes,
Ive methylated a ~300 kDa complex once, just for kicks and it still functioned. MS was fairly good as well, showing that essentially all accessible amines were altered. Notably I did not get the entire 300 kDa complex on MS it did not survive TFA/Acetonitrile and I saw components instead. There should not be a limit based on mass, as long as youre careful to maintain conditions that are both conducive to methylation and to the survival of your complex. Methylaiton can affect complex stability in either direction. Artem _____ From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of Jan Rash Sent: Tuesday, November 24, 2009 11:46 AM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Methylation of macromolecular complexes Hi all, What I really want to know is whether macromolecular complexes ( more than 200kDa) can tolerate methylation? thanks, Jan Rash 2009/11/24 Hüsnü Topal <huesnue.to...@rub.de> Hi Jan, Maybe this link <http://www.jenabioscience.com/cms/en/1/catalog/529_jbs_methylation_kit.html > http://www.jenabioscience.com/cms/en/1/catalog/529_jbs_methylation_kit.html or these references helps References [1] Kim et al. (2008) Large-scale evaluation of protein reductive methylation for improving protein crystallization. Nature Methods 5, 853. [2] Fogg et al. (2006) Application of the use of high-throughput technologies to the determination of protein structures of bacterial and viral pathogens. Acta Cryst. D62:1196. [3] Walter et al. (2006) Lysine methylation as a routine rescue strategy for protein crystallization. Structure 14:1617. Im not sure if this answers your question. Good luck. best regards, Hüsnü Topal Ruhr-Universität Bochum Institut für Physiologische Chemie, MA 2/141 Abteilung für Biochemie supramolekularer Systeme Universitätsstr. 150 44801 Bochum, Germany phone: +49 (0) 234 32 24934 Internet: <http://www.ruhr-uni-bochum.de/physiolchem/steegborn/> http://www.ruhr-uni-bochum.de/physiolchem/steegborn/ Diese E-Mail enthält vertrauliche und/oder rechtlich geschützte Informationen. Wenn Sie nicht der richtige Adressat sind oder diese E-Mail irrtümlich erhalten haben, informieren Sie bitte sofort den Absender und vernichten Sie diese Mail. Das unerlaubte Kopieren sowie die unbefugte Weitergabe dieser Mail ist nicht gestattet. This e-mail may contain confidential and/or privileged information. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden. J Go green Bitte denken Sie an die Umwelt und den Klimaschutz und drucken diese E-Mail nur aus, wenn unbedingt notwendig! Please consider the environment before printing this email! _____ Von: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] Im Auftrag von Jan Rash Gesendet: Dienstag, 24. November 2009 18:28 An: CCP4BB@JISCMAIL.AC.UK Betreff: [ccp4bb] Methylation of macromolecular complexes Hi all, I have a question regarding the methylation of macromolecular complexes. Is there any report where a macromolecular complex has been successfully methylated and later on crystallized. Is methylation a method of choice to chemically modify macromolecular complexes where the entropical contributions by lysines might be a cause for not obtaining crystals, or is there any other better ways? thanks, Jan Rash
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