A protein methylation pathway in Chlamydomonas flagella is active during flagellar resorption

Author Posting. © American Society for Cell Biology, 2008. This article is posted here by permission of American Society for Cell Biology for personal use, not for redistribution. The definitive version was published in Molecular Biology of the Cell 19 (2008): 4319-4327, doi:10.1091/mbc.E08-05-0470.During intraflagellar transport (IFT), the regulation of motor proteins, the loading and unloading of cargo and the turnover of flagellar proteins all occur at the flagellar tip. To begin an analysis of the protein composition of the flagellar tip, we used difference gel electrophoresis to compare long versus short (i.e., regenerating) flagella. The concentration of tip proteins should be higher relative to that of tubulin (which is constant per unit length of the flagellum) in short compared with long flagella. One protein we have identified is the cobalamin-independent form of methionine synthase (MetE). Antibodies to MetE label flagella in a punctate pattern reminiscent of IFT particle staining, and immunoblot analysis reveals that the amount of MetE in flagella is low in full-length flagella, increased in regenerating flagella, and highest in resorbing flagella. Four methylated proteins have been identified in resorbing flagella, using antibodies specific for asymmetrically dimethylated arginine residues. These proteins are found almost exclusively in the axonemal fraction, and the methylated forms of these proteins are essentially absent in full-length and regenerating flagella. Because most cells resorb cilia/flagella before cell division, these data indicate a link between flagellar protein methylation and progression through the cell cycle.This work was supported by National Institutes of Health Grant DK071720 (R.D.S.) and National Science Foundation Grant MCB 0418877 (R.D.S.)

Surface enhanced Raman spectroscopy on silver-nanoparticle-coated carbon-nanotube networks fabricated by electrophoretic deposition

In this study, the efficiency of silver nanoparticle (AgNP) decorated carbon nanotube (CNT) based porous substrates has been investigated for surface-enhanced Raman spectroscopy (SERS) applications. The fabrication of uniform thin coatings of carbon nanotubes is accomplished by Electrophoretic Deposition (EPD) on organosilane functionalized silicon substrates. The deposition process exemplifies a fast, reproducible and single-step room temperature coating strategy to fabricate horizontally aligned porous CNT network. Surfactant stabilized AgNPs were deposited on the CNT networks by immersion coating. The acquired Raman spectra of Rhodamine6G (R6G) analyte examined on the fabricated Ag-CNT-Si substrates exhibited enhanced signal intensity values when compared to SERS-active planar AgNP-Si substrates. An overall enhancement factor of ~;109 was achieved for the tested analyte which enables pushing the limit of detection to 1 × 10-12 M (1 pM). The enhancement can be attributed to the large surface area offered by the AgNP-CNT porous network, which is expected to increase the number of effective hot spots for the SERS effect. © 2014 The Korean Institute of Metals and Materials and Springer Science+Business Media Dordrecht