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Dsc protein complex sequence alignments.
We evaluated the binding of purified AafA- dsc protein to integrin α5 β1 and fibronectin/integrin α5 β1 complex.
No direct binding of AafA- dsc protein to integrin α5 β1 was found; however, a significant binding to fibronectin/integrin α5 β1 complex was observed).
Similarly, we found that AafA- dsc protein binds to fibronectin/integrin α5 β1 complex but not to integrin α5 β1 alone.
The latter observation is supported by coimmunoprecipitation assays using purified AafA- dsc protein that confirmed the formation of the complex AafA/fibronectin/integrin α5 β1.
To evaluate the formation of the AafA/fibronectin complex, we prepared a solution of 5 μg/mL of AafA- dsc protein and 10 μg/mL of purified fibronectin.
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"Our studies suggest that dsc proteins are an attractive target for the development of new drugs to combat fungal infections in immune-compromised people," Espenshade said.
The Dsc proteins form a complex in the Golgi membrane that resembles E3 ligases involved in ER-associated protein degradation [ 32].
For DSC analysis, proteins were diluted to a concentration of 5 μM in PBS, pH 6.0 or 7.4, followed by degassing and analysis by using a VP-DSC Capillary Cell MicroCalorimeter (MicroCal, Northampton, MA).
This approach is interrogation of DSCs for proteins and other labile cellular factors that are detected specifically on either immortal chromosomes or mortal chromosomes.
Furthermore, the basal level of cytoplasmic L-pSMAD1 signal observed in cells under BMP and control conditions suggest a continuous turn-over of the Smad1 protein in DSC neural precursor cells.
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