WikiSysop: Created page with "== Citation == Levitz, Talya S. / Brignole, Edward J. / Fong, Ivan / Darrow, Michele C. / Drennan, Catherine L. Effects of chameleon dispense-to-plunge speed on particle conc..."

2022-06-09T14:33:14Z

Created page with "== Citation == Levitz, Talya S. / Brignole, Edward J. / Fong, Ivan / Darrow, Michele C. / Drennan, Catherine L. Effects of chameleon dispense-to-plunge speed on particle conc..."

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== Citation ==

Levitz, Talya S. / Brignole, Edward J. / Fong, Ivan / Darrow, Michele C. / Drennan, Catherine L. Effects of chameleon dispense-to-plunge speed on particle concentration, complex formation, and final resolution: A case study using the Neisseria gonorrhoeae ribonucleotide reductase inactive complex. 2022. Journal of structural biology, Vol. 214, p. 107825

== Abstract ==

Ribonucleotide reductase (RNR) is an essential enzyme that converts ribonucleotides to deoxyribonucleotides and is a promising antibiotic target, but few RNRs have been structurally characterized. We present the use of the chameleon, a commercially-available piezoelectric cryogenic electron microscopy plunger, to address complex denaturation in the Neisseria gonorrhoeae class Ia RNR. Here, we characterize the extent of denaturation of the ring-shaped complex following grid preparation using a traditional plunger and using a chameleon with varying dispense-to-plunge times. We also characterize how dispense-to-plunge time influences the amount of protein sample required for grid preparation and preferred orientation of the sample. We demonstrate that the fastest dispense-to-plunge time of 54 ms is sufficient for generation of a data set that produces a high quality structure, and that a traditional plunging technique or slow chameleon dispense-to-plunge times generate data sets limited in resolution by complex denaturation. The 4.3 Å resolution structure of Neisseria gonorrhoeae class Ia RNR in the inactive α4β4 oligomeric state solved using the chameleon with a fast dispense-to-plunge time yields molecular information regarding similarities and differences to the well studied Escherichia coli class Ia RNR α4β4 ring.

== Keywords ==

== Links ==

https://www.sciencedirect.com/science/article/pii/S1047847721001301

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== Comments ==

2022Levitz Chameleon - Revision history

WikiSysop: Created page with "== Citation == Levitz, Talya S. / Brignole, Edward J. / Fong, Ivan / Darrow, Michele C. / Drennan, Catherine L. Effects of chameleon dispense-to-plunge speed on particle conc..."