PubMed Health⌕ Search
Home/Archive/PubMed

PubMed · 8109963

DESIGN: a guide to protein crystallization experiments.

J Sedzik
View original source ↗Export metadata ↓

Abstract

Existing guidelines for crystallizing proteins generally require substantial amounts of protein and scarce laboratory resources. To address this problem we have developed the program DESIGN, which, suited to daily laboratory usage, utilizes statistics when screening variables of crystallization. The program DESIGN is described in considerable detail and examples of its outcome and capabilities are shown. In addition, a new scoring scale for crystallization trials is described. It is expected that the use of such an approach may provide additional constraints toward crystallizing the protein, even if the conditions applied are not successful.

Explore related subjects

CalmodulinCations, DivalentConcanavalin ACrystallizationCrystallography, X-RayIndicators and ReagentsLanthanumMuramidaseMyelin ProteinsProteinsSoftwareJournal ArticleResearch Support, U.S. Gov't, P.H.S.

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

J Sedzik. 1994-02-01. DESIGN: a guide to protein crystallization experiments.. https://doi.org/10.1006/abbi.1994.1049

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

KEEP EXPLORING

Related citations

PubMed2026

Cryo-EM structure of soluble VPS13C suggests its regulation by a conformational switch and by calmodulin.

Bridge-like lipid transfer proteins (BLTPs) play fundamental roles in cellular lipid redistribution between organellar membranes. They comprise bridge domains spanning organelles at contact sites that allow lipids to transit through the cytosol between adjacent membranes. The assembly of BLTPs into complexes with adaptor proteins enables lipid transfer. To address the mechanisms underlying the assembly and regulation of BLTP complexes, we used cryo-EM to resolve the structure of one such BLTP, the Parkinson's disease protein VPS13C, at near-atomic resolution. The structure identifies a lipid-transfer-nonpermissive conformation, in which the built-in C-terminal VAB adaptor module blocks the end of the lipid transfer bridge, interfering with lipid delivery. We also identify calmodulin (CaM), central to calcium signaling, as a constitutive VPS13C interactor. Calcium induces conformational changes in the VPS13C-CaM complex, suggesting calcium regulation of VPS13 function. Altogether, this structure of intact VPS13C serves as a starting point for understanding its regulation and that of other VPS13 proteins.

Calmodulin↗
PubMed2025

Calmodulin D133H Disrupts Cav1.2 and Kv7.1 Regulation to Prolong Cardiac Action Potentials in Long QT Syndrome.

Calmodulin (CaM) plays a central role in cardiac excitation-contraction coupling by regulating ion channels, including the L-type calcium (Ca2+) channel Cav1.2 and the voltage-gated potassium (K+) channel Kv7.1. Mutations in CaM are linked to severe arrhythmogenic disorders such as Long QT syndrome (LQTS), yet the molecular mechanisms remain incompletely understood. Here, we investigate the structural and functional consequences of the arrhythmia-associated CaM variant D133H. Biophysical analysis revealed that D133H destabilises Ca2+ binding at the C-terminal lobe of CaM, altering its Ca2+-dependent conformational changes. Electrophysiological recordings demonstrated that CaM D133H impairs Ca2+-dependent inactivation (CDI) of Cav1.2, prolonging Ca2+ influx, while also reducing activation of Kv7.1, thereby limiting repolarising K+ currents. Together, these dual defects converge to prolong action potential duration, providing a mechanistic basis for arrhythmogenesis in LQTS. Our findings establish that CaM D133H perturbs both Ca2+ and K+ channel regulation, highlighting a shared pathway by which calmodulinopathy mutations disrupt cardiac excitability.

Calmodulin↗