Cyrus Tang Hematology Center, MOE Engineering Center of Hematological Disease, Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.
MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China.
Sci Rep. 2017 Mar 24;7:45262. doi: 10.1038/srep45262.
Membrane-bound proteases are essential for epidermal integrity. Human airway trypsin-like protease 4 (HAT-L4) is a type II transmembrane serine protease. Currently, its biochemical property, cellular distribution and physiological function remain unknown. Here we examined HAT-L4 expression and function in vitro and in vivo. In Western analysis, HAT-L4 expressed in transfected CHO cells appeared as a 48-kDa protein. Flow cytometry confirmed HAT-L4 expression on the cell surface with the expected membrane topology. RT-PCR and immunostaining experiments indicated that HAT-L4 was expressed in epithelial cells and exocrine glands in tissues including skin, esophagus, trachea, tongue, eye, bladder, testis and uterus. In the skin, HAT-L4 expression was abundant in keratinocytes and sebaceous glands. We generated HAT-L4 knockout mice by disrupting the Tmprss11f gene encoding HAT-L4. HAT-L4 knockout mice were viable and fertile. No defects were found in HAT-L4 knockout mice in hair growth, wound healing, water repulsion and body temperature regulation. Compared with wild-type controls, HAT-L4-deficient newborn mice had greater body fluid loss and higher mortality in a trans-epidermal body fluid loss test. In metabolic studies, HAT-L4-deficient adult mice drank water more frequently than wild-type controls did. These results indicate that HAT-L4 is important in epidermal barrier function to prevent body fluid loss.
膜结合蛋白酶对于表皮完整性至关重要。人类气道胰蛋白酶样蛋白酶 4(HAT-L4)是一种 II 型跨膜丝氨酸蛋白酶。目前,其生化特性、细胞分布和生理功能尚不清楚。在这里,我们研究了 HAT-L4 在体外和体内的表达和功能。在 Western blot 分析中,转染 CHO 细胞中表达的 HAT-L4 表现为 48kDa 的蛋白。流式细胞术证实 HAT-L4 在细胞表面表达,具有预期的膜拓扑结构。RT-PCR 和免疫染色实验表明,HAT-L4 在皮肤、食管、气管、舌、眼、膀胱、睾丸和子宫等组织的上皮细胞和外分泌腺中表达。在皮肤中,HAT-L4 在角质形成细胞和皮脂腺中表达丰富。我们通过破坏编码 HAT-L4 的 Tmprss11f 基因,生成了 HAT-L4 敲除小鼠。HAT-L4 敲除小鼠具有活力和生育能力。在毛发生长、伤口愈合、水排斥和体温调节方面,HAT-L4 敲除小鼠没有发现缺陷。与野生型对照相比,HAT-L4 缺陷型新生小鼠在经表皮体液丢失试验中具有更大的体液丢失和更高的死亡率。在代谢研究中,HAT-L4 缺陷型成年小鼠比野生型对照更频繁地饮水。这些结果表明 HAT-L4 对于表皮屏障功能很重要,可防止体液丢失。