Harvard Medical School, Boston, MA, USA.
Am J Physiol Cell Physiol. 2010 Jun;298(6):C1326-42. doi: 10.1152/ajpcell.00552.2009. Epub 2010 Feb 24.
Proton-transporting cells are located in several tissues where they acidify the extracellular environment. These cells express the vacuolar H(+)-ATPase (V-ATPase) B1 subunit (ATP6V1B1) in their plasma membrane. We provide here a comprehensive catalog of the proteins that are expressed in these cells, after their isolation by enzymatic digestion and fluorescence-activated cell sorting (FACS) from transgenic B1-enhanced green fluorescent protein (EGFP) mice. In these mice, type A and B intercalated cells and connecting segment cells of the kidney, and narrow and clear cells of the epididymis, which all express ATP6V1B1, also express EGFP, while all other cell types are negative. The proteome of renal and epididymal EGFP-positive (EGFP(+)) cells was identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and compared with their respective EGFP-negative (EGFP(-)) cell populations. A total of 2,297 and 1,564 proteins were detected in EGFP(+) cells from the kidney and epididymis, respectively. Out of these proteins, 202 and 178 were enriched by a factor greater than 1.5 in EGFP(+) cells compared with EGFP(-) cells, in the kidney and epididymis respectively, and included subunits of the V-ATPase (B1, a4, and A). In addition, several proteins involved in intracellular trafficking, signaling, and cytoskeletal dynamics were identified. A novel common protein that was enriched in renal and epididymal EGFP(+) cells is the progesterone receptor, which might be a potential candidate for the regulation of V-ATPase-dependent proton transport. These proteomic databases provide a framework for comprehensive future analysis of the common and distinct functions of V-ATPase-B1-expressing cells in the kidney and epididymis.
质子转运细胞位于几种组织中，在这些组织中，它们使细胞外环境酸化。这些细胞在质膜中表达液泡 H(+)-ATP 酶 (V-ATPase) B1 亚基 (ATP6V1B1)。在这里，我们提供了一个综合目录，其中列出了通过从转基因 B1-增强型绿色荧光蛋白 (EGFP) 小鼠中通过酶消化和荧光激活细胞分选 (FACS) 分离这些细胞后，这些细胞中表达的蛋白质。在这些小鼠中，A 型和 B 型闰细胞和连接段细胞、肾脏的和窄而透明细胞的附睾，所有这些细胞都表达 ATP6V1B1，也表达 EGFP，而所有其他细胞类型均为阴性。通过液相色谱-串联质谱法 (LC-MS/MS) 鉴定了肾脏和附睾 EGFP 阳性 (EGFP(+)) 细胞的蛋白质组，并将其与各自的 EGFP 阴性 (EGFP(-)) 细胞群体进行了比较。在肾脏和附睾的 EGFP(+)细胞中分别检测到 2,297 和 1,564 种蛋白质。在这些蛋白质中，在肾脏和附睾中，分别有 202 和 178 种蛋白质在 EGFP(+)细胞中比 EGFP(-)细胞中富集超过 1.5 倍，包括 V-ATPase (B1、a4 和 A) 的亚基。此外，还鉴定了几种参与细胞内运输、信号转导和细胞骨架动力学的蛋白质。一种在肾脏和附睾的 EGFP(+)细胞中富集的新型共同蛋白质是孕激素受体，它可能是调节 V-ATPase 依赖性质子转运的潜在候选物。这些蛋白质组数据库为全面分析肾脏和附睾中 V-ATPase-B1 表达细胞的共同和独特功能提供了框架。
