Several peripheral membrane proteins are known to form membrane pores through multimerization. In many cases, in biochemical reconstitution experiments, a complex distribution of oligomeric states has been observed that may, in part, be irrelevant to their physiological functions. This phenomenon makes it difficult to identify the functional oligomeric states of membrane lipid interacting proteins, for example, during the formation of transient membrane pores. Using fibroblast growth factor 2 (FGF2) as an example, we present a methodology applicable to giant lipid vesicles by which functional oligomers can be distinguished from nonspecifically aggregated proteins without functionality. Two distinct populations of fibroblast growth factor 2 were identified with (i) dimers to hexamers and (ii) a broad population of higher oligomeric states of membrane-associated FGF2 oligomers significantly distorting the original unfiltered histogram of all detectable oligomeric species of FGF2. The presented statistical approach is relevant for various techniques for characterizing membrane-dependent protein oligomerization.
Neuroblastoma cell line SH-SY5Y, due to its capacity to differentiate into neurons, easy handling, and low cost, is a common experimental model to study molecular events leading to Alzheimer's disease (AD). However, it is prevalently used in its undifferentiated state, which does not resemble neurons affected by the disease. Here, we show that the expression and localization of amyloid-β protein precursor (AβPP), one of the key molecules involved in AD pathogenesis, is dramatically altered in SH-SY5Y cells fully differentiated by combined treatment with retinoic acid and BDNF. We show that insufficient differentiation of SH-SY5Y cells results in AβPP mislocalization.
- MeSH
- Alzheimerova nemoc metabolismus MeSH
- amyloidový prekurzorový protein beta metabolismus MeSH
- biologické modely MeSH
- buněčná diferenciace fyziologie MeSH
- intravitální mikroskopie metody MeSH
- lidé MeSH
- mozkový neurotrofický faktor * metabolismus farmakologie MeSH
- nádorové buněčné linie MeSH
- neuroblastom MeSH
- neurony fyziologie MeSH
- oxidační stres MeSH
- proteolýza MeSH
- tretinoin * metabolismus farmakologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- dopisy MeSH