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153.
  • Jafferali, Mohammed Hakim, et al. (creator_code:aut_t)
  • MCLIP, an effective method to detect interactions of transmembrane proteins of the nuclear envelope in live cells
  • 2014
  • record:In_t: Biochimica et Biophysica Acta - Biomembranes. - : Elsevier BV. - 0005-2736 .- 1879-2642 .- 0006-3002. ; 1838:10, s. 2399-2403
  • swepub:Mat_article_t (swepub:level_refereed_t)abstract
    • Investigating interactions of proteins in the nuclear envelope (NE) using co-immunoprecipitation (Co-IP) has previously been difficult or even impossible due to their inherent resistance to extraction. We have developed a novel method, MCLIP (Membrane protein Cross-Link ImmunoPrecipitation), which takes advantage of a cell permeable crosslinker to enable effective detection and analysis of specific interactions of NE proteins in live cells using Western blot. Using MCLIP we show that, in U2OS cells, the integral inner nuclear membrane protein Samp1 interacts with Lamin B1, the LINC (Linker of nucleoskeleton and cytoskeleton) complex protein, Sun1 and the soluble small GTPase Ran. The results show that the previously detected in vitro interaction between Samp1 and Emerin also takes place in live cells. In vitro pull down experiments show, that the nucleoplasmic domains of Samp1 and Emerin can bind directly to each other. We also, show that MCLIP is suitable to coprecipitate protein interactions in different stages of the cell cycle.
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154.
  • Kohli, Gurjeet S, et al. (creator_code:aut_t)
  • Polyketide synthesis genes associated with toxin production in two species of Gambierdiscus (Dinophyceae).
  • 2015
  • record:In_t: BMC Genomics. - : Springer Science and Business Media LLC. - 1471-2164. ; 16
  • swepub:Mat_article_t (swepub:level_refereed_t)abstract
    • Marine microbial protists, in particular, dinoflagellates, produce polyketide toxins with ecosystem-wide and human health impacts. Species of Gambierdiscus produce the polyether ladder compounds ciguatoxins and maitotoxins, which can lead to ciguatera fish poisoning, a serious human illness associated with reef fish consumption. Genes associated with the biosynthesis of polyether ladder compounds are yet to be elucidated, however, stable isotope feeding studies of such compounds consistently support their polyketide origin indicating that polyketide synthases are involved in their biosynthesis.
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155.
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156.
  • Larsson, Veronica J., et al. (creator_code:aut_t)
  • Mitotic spindle assembly and γ-tubulin localisation depend on the integral nuclear membrane protein, Samp1
  • 2018
  • record:In_t: Journal of Cell Science. - : The Company of Biologists. - 0021-9533 .- 1477-9137. ; 131:8
  • swepub:Mat_article_t (swepub:level_refereed_t)abstract
    • We have investigated a possible role of the inner nuclear membrane protein, Samp1, in the mitotic machinery. Live cell imaging showed that Samp1aYFP distributed as filamentous structures in the mitotic spindle, partially co-localising with ß-tubulin. Samp1 depletion resulted in an increased frequency of cells with signs of chromosomal mis-segregation and prolonged metaphase, indicating problems with spindle assembly and/or chromosomal alignment. Consistently, mitotic spindles in Samp1 depleted cells contained significantly lower levels of ß-tubulin and γ-tubulin, phenotypes which were rescued by overexpression of Samp1aYFP. We found that Samp1 can bind directly to γ-tubulin and that Samp1 co-precipitated with γ-tubulin and HAUS6 of the Augmin complex in live cells. The levels of Haus6, in the mitotic spindle also decreased after Samp1 depletion. We show that Samp1 is involved in the recruitment of Haus6 and γ-tubulin to the mitotic spindle. Samp1 is the first inner nuclear membrane protein shown to have a function in mitotic spindle assembly.
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159.
  • Lundblad, Lennart, et al. (creator_code:aut_t)
  • Thoracic gas volume measurements in paralyzed mice
  • 2004
  • record:In_t: Annals of Biomedical Engineering. - 1573-9686. ; 32:10, s. 1420-1427
  • swepub:Mat_article_t (swepub:level_refereed_t)abstract
    • We have previously measured thoracic gas volume (V-TG) in spontaneously breathing mice using a whole body plethysmograph and have now extended our technique to allow for V-TG measurements during paralysis. BALB/c mice were anesthetized and placed in a body-box and ventilated via a tracheostomy cannula through the box wall. Box pressure (P-b) and tracheal pressure (P-ao) were measured during spontaneous breathing, and again after paralysis while mechanically compressing the chest. V-TG was much larger after paralysis (0.49+/-0.06 ml, positive end-expiratory pressure=2 cmH(2)O) when compared with spontaneous breathing (0.31+/-0.01 ml). External chest compression produced looping in the plots of P-b versus P-ao that was attributable to gradual changes in P-b upon release of the mechanical chest compression and had the character of thermal transients. Under the assumption that the rate of heating of the air in the chamber was proportional to the pressure applied to the animal's chest, and that any increase in air temperature was dissipated by heat absorption by the chamber walls, we developed an algorithm that corrected for the thermal events. This yielded similar results for V-TG (0.30+/-0.02 ml) as obtained during spontaneous efforts. Our method may prove particularly useful when paralysis is required for the precise measurement of lung mechanics.
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