MCC/eisosomes are protein-organized domains in the plasma membrane of fungi and algae. However, the composition and function(s) of MCC/eisosomes in the filamentous fungus Neurospora crassa were previously unknown. To identify proteins that localize to MCC/eisosomes in N. crassa, we isolated proteins that co-purified with the core MCC/eisosome protein LSP-1, which was tagged with GFP. Proteins that co-fractionated with LSP-1:GFP were then identified by mass spectrometry. Eighteen proteins were GFP-tagged and used to identify six proteins that highly colocalized with the MCC/eisosome marker LSP-1:RFP, while five other proteins showed partial overlap with MCC/eisosomes. Seven of these proteins showed amino acid sequence homology with proteins known to localize to MCC/eisosomes in the yeast Saccharomyces cerevisiae. However, homologs of three proteins known to localize to MCC/eisosomes in S. cerevisiae (Can1, Pkh1/2, and Fhn1) were not found to colocalize with MCC/eisosome proteins in N. crassa by fluorescence microscopy. Interestingly, one new eisosome protein (glutamine-fructose-6-phosphate aminotransferase, gene ID: NCU07366) was detected in our studies. These findings demonstrate that there are interspecies differences of the protein composition of MCC/eisosomes. To gain further insight, molecular modeling and bioinformatics analysis of the identified proteins were used to propose the organization of MCC/eisosomes in N. crassa. A model will be discussed for how the broad range of functions predicted for the proteins localized to MCC/eisosomes, including cell wall synthesis, response and signaling, transmembrane transport, and actin organization, suggests that MCC/eisosomes act as organizing centers in the plasma membrane.

MCC /异构体是真菌和藻类质膜中的蛋白质组织结构域。然而，丝状真菌中MCC /同质体的组成和功能克氏菌以前是未知的。识别位于MCC /同质异体中的蛋白质克拉萨猪笼草，我们分离了与核心MCC /同质体蛋白LSP-1共纯化的蛋白，该蛋白用GFP标记。然后通过质谱鉴定与LSP-1：GFP共分离的蛋白质。18个蛋白经GFP标记，用于鉴定与MCC /酶体标记LSP-1：RFP高度共定位的6个蛋白，而其他5个蛋白与MCC /酶体部分重叠。这些蛋白质中的七个显示出与已知位于酵母中MCC /异构体的蛋白质的氨基酸序列同源性酿酒酵母。但是，已知三种蛋白质的同源物位于MCC /等位体中酿酒酵母 （Can1，Pkh1 / 2，和Fhn1）未发现与MCC /同质体蛋白共定位 克拉萨猪笼草通过荧光显微镜。有趣的是，在我们的研究中检测到一种新的酶体蛋白（谷氨酰胺-果糖-6-磷酸氨基转移酶，基因ID：NCU07366）。这些发现表明，MCC /异构体的蛋白质组成存在种间差异。为了获得更多的见解，对鉴定出的蛋白质进行了分子建模和生物信息学分析，以提出MCC /同质体的组织。克拉萨猪笼草。将讨论一个模型，该模型预测了定位于MCC /异构体的蛋白质的广泛功能，包括细胞壁合成，应答和信号传导，跨膜转运和肌动蛋白组织，表明MCC /异构体在血浆中起组织中心作用膜。