Coiled Coils Direct Assembly of a Cold-Activated TRP Channel

SummaryTransient receptor potential (TRP) channels mediate numerous sensory transduction processes and are thought to function as tetramers. TRP channel physiology is well studied; however, comparatively little is understood regarding TRP channel assembly. Here, we identify an autonomously folded assembly domain from the cold- and menthol-gated channel TRPM8. We show that the TRPM8 cytoplasmic C-terminal domain contains a coiled coil that is necessary for channel assembly and sufficient for tetramer formation. Cell biological experiments indicate that coiled-coil formation is required for proper channel maturation and trafficking and that the coiled-coil domain alone can act as a dominant-negative inhibitor of functional channel expression. Our data define an authentic TRP modular assembly domain, establish a clear role for coiled coils in ion channel assembly, demonstrate that coiled-coil assembly domains are a general feature of TRPM channels, and delineate a new tool that should be of general use in dissecting TRPM channel function

Comparison of the Hindin Proteins of Strongylocentrotus franciscanus, S. purpuratus, and Lytechinus variegatus: Sequences involved in the Species Specificity of Fertilization

Bindin is the sea urchin sperm acrosomal protein that is responsible for the species-specific adhesion of the sperm to the egg. Two new bindin cDNA sequences that contain the entire open reading frame for the binding precursor are reported: one for Strongylocentrotus franciscanus and one for Lytechinus variegatus. Both contain inverted repetitive sequences in their 3' untranslated regions, and the S. franciscanus cDNA contains an inverted repetitive sequence match between the 5' untranslated region and the coding region. The middle third of the mature bindin sequence is highly conserved in all three species, and the flanking sequences share short repeated sequences that vary in number between the species. Cross-fertilization data are reported for the species S. purpuratus, S. franciscanus, L. variegatus, and L. pictus. A barrier to cross-fertilization exists between the sympatric Strongylocentrotus species, but there is no barrier between the allopatric Lytechinus species

Nanoscale mosaicity revealed in peptide microcrystals by scanning electron nanodiffraction.

Changes in lattice structure across sub-regions of protein crystals are challenging to assess when relying on whole crystal measurements. Because of this difficulty, macromolecular structure determination from protein micro and nanocrystals requires assumptions of bulk crystallinity and domain block substructure. Here we map lattice structure across micron size areas of cryogenically preserved three-dimensional peptide crystals using a nano-focused electron beam. This approach produces diffraction from as few as 1500 molecules in a crystal, is sensitive to crystal thickness and three-dimensional lattice orientation. Real-space maps reconstructed from unsupervised classification of diffraction patterns across a crystal reveal regions of crystal order/disorder and three-dimensional lattice tilts on the sub-100nm scale. The nanoscale lattice reorientation observed in the micron-sized peptide crystal lattices studied here provides a direct view of their plasticity. Knowledge of these features facilitates an improved understanding of peptide assemblies that could aid in the determination of structures from nano- and microcrystals by single or serial crystal electron diffraction

CMM : an enhanced platform for interactive validation of metal binding sites

Abstract Metal ions bound to macromolecules play an integral role in many cellular processes. They can directly participate in catalytic mechanisms or be essential for the structural integrity of proteins and nucleic acids. However, their unique nature in macromolecules can make them difficult to model and refine, and a substantial portion of metal ions in the PDB are misidentified or poorly refined. CheckMyMetal (CMM) is a validation tool that has gained widespread acceptance as an essential tool for researchers working on metal-macromolecule complexes. CMM can be used during structure determination or to validate metal binding sites in structural models within the PDB. The functionalities of CMM have recently been greatly enhanced and provide researchers with additional information that can guide modeling decisions. The new version of CMM shows metals in the context of electron density maps and allows for on-the-fly refinement of metal binding sites. The improvements should increase the reproducibility of biomedical research. The web server is available at https://cmm.minorlab.org

Absorption Spectral Slopes and Slope Ratios as Indicators of Molecular Weight, Source, and Photobleaching of Chromophoric Dissolved Organic Matter

A new approach for parameterizing dissolved organic matter ( DOM) ultraviolet-visible absorption spectra is presented. Two distinct spectral slope regions ( 275-295 nm and 350-400 nm) within log-transformed absorption spectra were used to compare DOM from contrasting water types, ranging from wetlands (Great Dismal Swamp and Suwannee River) to photobleached oceanic water ( Atlantic Ocean). On the basis of DOM size-fractionation studies ( ultrafiltration and gel filtration chromatography), the slope of the 275-295- nm region and the ratio of these slopes (SR; 275-295- nm slope : 350-400- nm slope) were related to DOM molecular weight ( MW) and to photochemically induced shifts in MW. Dark aerobic microbial alteration of chromophoric DOM ( CDOM) resulted in spectral slope changes opposite of those caused by photochemistry. Along an axial transect in the Delaware Estuary, large variations in SR were measured, probably due to mixing, photodegradation, and microbial alteration of CDOM as terrestrially derived DOM transited through the estuary. Further, SR varied by over a factor of 13 between DOM-rich wetland waters and Sargasso Sea surface waters. Currently, there is no consensus on a wavelength range for log-transformed absorption spectra. We propose that the 275-295- nm slope be routinely reported in future DOM studies, as it can be measured with high precision, it facilitates comparison among dissimilar water types including CDOM-rich wetland and CDOM-poor marine waters, and it appears to be a good proxy for DOM MW. © 2008, by the American Society of Limnology and Oceanography, Inc

Corn, 1992

Harry C. Minor is an Associate Professor of Agronomy and State Extension Specialist, Carl G. Morris and Howard L. Mason are Senior Research Specialists, and Delbert R. Knerr, David B. Thomas and C. Stephen Lankheit are Research Specialists in Agronomy.Compares hybrids and includes experimental procedures, yield results and seed corn company addresses.Comparing hybrids -- Experimental procedures -- Summary of results -- Yield results -- Seed corn company addresse