Search PubMed⌕ Search
Home/Archive/PubMed

PubMed · 12351856

Screening crystallisation conditions using fluorescence correlation spectroscopy.

Ralf Schmauder·Thomas Schmidt·Jan Pieter Abrahams·Maxim E Kuil
View original source ↗Export metadata ↓

Abstract

We investigate the potential of fluorescence correlation spectroscopy (FCS) in screening for crystallisation conditions. Solutions that nucleate protein crystals must have different interactions than solutions that do not give rise to crystals. Due to these different interactions the average mean squared displacement of the individual proteins changes. By monitoring protein self-diffusion, we can distinguish crystallizing from non-crystallizing solutions. The method introduced can be applied at extremely low concentrations in femtoliter volumes as an early diagnostic for molecular association. Based on our preliminary findings FCS has the potential to become a routine screening method for crystallography.

Explore related subjects

ApoferritinsCarbocyaninesCrystallizationDiffusionFluorescent DyesMuramidaseProteinsSolutionsSpectrometry, FluorescenceJournal ArticleResearch Support, Non-U.S. Gov't

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Ralf Schmauder, Thomas Schmidt, Jan Pieter Abrahams, Maxim E Kuil. 2002-09-26. Screening crystallisation conditions using fluorescence correlation spectroscopy.. https://doi.org/10.1107/s0907444902014373

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related citations

PubMed2006

Structure of dual function iron regulatory protein 1 complexed with ferritin IRE-RNA.

Iron regulatory protein 1 (IRP1) binds iron-responsive elements (IREs) in messenger RNAs (mRNAs), to repress translation or degradation, or binds an iron-sulfur cluster, to become a cytosolic aconitase enzyme. The 2.8 angstrom resolution crystal structure of the IRP1:ferritin H IRE complex shows an open protein conformation compared with that of cytosolic aconitase. The extended, L-shaped IRP1 molecule embraces the IRE stem-loop through interactions at two sites separated by approximately 30 angstroms, each involving about a dozen protein:RNA bonds. Extensive conformational changes related to binding the IRE or an iron-sulfur cluster explain the alternate functions of IRP1 as an mRNA regulator or enzyme.

Apoferritins↗
PubMed2006

Versatile apoferritin nanoparticle labels for assay of protein.

A versatile bioassay label based on marker-loaded apoferritin nanoparticles (MLANs) has been developed for sensitive protein detection. Dissociation and reconstitution characteristics at different pH as well as the special cavity structure of apoferritin provides a facile route to prepare nanoparticle labels and avoid the complicated and tedious synthesis process of conventional nanoparticle labels. The optical and electrochemical characteristics of the prepared nanoparticle labels are easily controlled by loading different optical or electrochemical markers. A fluorescence marker (fluorescein anion) and a redox marker [hexacyanoferrate(III)] were used as model markers to load into the cavity of apoferritin nanoparticles for microscopic fluorescence immunoassay and electrochemical immunoassay, respectively. Detection limits of 0.06 (0.39 pM) and 0.08 ng mL(-1) (0.52 pM) IgG were obtained with fluorescein MLAN and hexacyanoferrate MLANs, respectively. The new nanoparticle labels hold great promise for multiplex protein detection (in connection with nanoparticles loaded with different markers) and for enhancing the sensitivity of other bioassays.

Apoferritins↗