expression-purification-crystallization-enzyme-assays

Expression, Purification, Crystallization, and Enzyme Assays of Fumarylacetoacetate Hydrolase Domain-Containing Proteins

Fumarylacetoacetate hydrolase (FAH) domain-containing proteins (FAHD) are identified members of the FAH superfamily in eukaryotes. Enzymes of this ...

Medical University of Innsbruck Austria

Reviewing biomolecular crystallography proposals: time for a paradigm change.

Afamin, a human plasma glycoprotein and putative transporter of hydrophobic molecules, has been shown to act as extracellular chaperone for poorly soluble, acylated Wnt proteins, forming a stable, soluble complex with functioning Wnt proteins. The 2.1-Å crystal structure of glycosylated human afamin reveals an almost exclusively hydrophobic binding cleft capable of harboring large hydrophobic moieties. Lipid analysis confirms the presence of lipids, and density in the primary binding pocket of afamin was modeled as palmitoleic acid, presenting the native O-acylation on serine 209 in human Wnt3a. The modeled complex between the experimental afamin structure and a Wnt3a homology model based on the XWnt8-Fz8-CRD fragment complex crystal structure is compelling, with favorable interactions comparable with the crystal structure complex. Afamin readily accommodates the conserved palmitoylated serine 209 of Wnt3a, providing a structural basis how afamin solubilizes hydrophobic and poorly soluble Wnt proteins.

Structural Evidence for a Role of the Multi-functional Human Glycoprotein Afamin in Wnt Transport.

The almost universally required "Table 1," summarizing data-collection and data-processing statistics, has in its present form outlived its usefulness in almost all publications of biomolecular crystal structure reports. Information contained in "Table 1" is insufficient to evaluate or repeat the experiment; is redundant with information extractable from deposited diffraction data; and includes data items whose meaning is under increased scrutiny in the crystallographic community. Direct and consistent extraction and analysis of data quality metrics from preferably unmerged intensity data with graphical presentation of reciprocal space features, including impact on map and model features, should replace "Table 1."

Article	Year
Reviewing biomolecular crystallography proposals: time for a paradigm change. Trends in biochemical sciences\| PubMed ID: 26051321	2015
Structural Evidence for a Role of the Multi-functional Human Glycoprotein Afamin in Wnt Transport. Structure (London, England : 1993)\| PubMed ID: 29153507	2017
Against Method: Table 1-Cui Bono? Structure (London, England : 1993)\| PubMed ID: 29861344	2018

Bernhard Rupp

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