Differences in proteasome targeting in Arabidopsis leaf mesophyll, epidermal and vascular cells

For the detailed and sensitive detection of the responses to a specific treatment it is important to perform tissue or cell type specific analyses, but this is not easily achievable for the different leaf tissues. Here we developed a method termed MeSelect to effectively separate leaf epidermis, vascular and mesophyll cells basically without contamination from other tissues. The high yield of the MeSelect method allowed for tissue specific high-throughput proteome analyses after inhibition of the proteasome with Syringolin A and the affinity enrichment of polyubiquitylated proteins in epidermis, mesophyll and vasculature.

为实现对特定处理响应的精准且灵敏的检测，开展组织或细胞类型特异性分析至关重要，但针对不同叶片组织开展此类分析并非易事。为此，我们开发了一种命名为MeSelect的方法，可有效分离叶片表皮、维管组织与叶肉细胞，且基本无其他组织污染。得益于MeSelect方法的高得率，我们得以在采用丁香菌素A（Syringolin A）抑制蛋白酶体后，针对表皮、叶肉与维管组织开展组织特异性高通量蛋白质组分析，并对这些组织中的多泛素化蛋白进行亲和富集。