State Key Laboratory of Medical Genomics, Research Center for Experimental Medicine, Rui-Jin Hospital affiliated to Shanghai Jiao Tong University School of Medicine, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200025, China.
Reproductive Medical Center, Rui-Jin Hospital affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
J Cell Sci. 2021 May 15;134(10). doi: 10.1242/jcs.258426. Epub 2021 May 24.
A disintegrin and metalloproteinase 3 (ADAM3) is a sperm membrane protein critical for sperm migration from the uterus into the oviduct and sperm-egg binding in mice. Disruption of PRSS37 results in male infertility concurrent with the absence of mature ADAM3 from cauda epididymal sperm. However, how PRSS37 modulates ADAM3 maturation remains largely unclear. Here, we determine the PRSS37 interactome by GFP immunoprecipitation coupled with mass spectrometry in PRSS37-EGFP knock-in mice. Three molecular chaperones (CLGN, CALR3 and PDILT) and three ADAM proteins (ADAM2, ADAM6B and ADAM4) were identified to be interacting with PRSS37. Coincidently, five of them (except ADAM4) have been reported to interact with ADAM3 precursor and regulate its maturation. We further demonstrated that PRSS37 also interacts directly with ADAM3 precursor and its deficiency impedes the association between PDILT and ADAM3. This could contribute to improper translocation of ADAM3 to the germ cell surface, leading to ADAM3 loss in PRSS37-null mature sperm. The understanding of the maturation mechanisms of pivotal sperm plasma membrane proteins will pave the way toward novel strategies for contraception and the treatment of unexplained male infertility.
解整合素金属蛋白酶 3（ADAM3）是一种精子膜蛋白，对于精子从子宫迁移到输卵管以及精子与卵子结合至关重要。PRSS37 的破坏导致雄性不育，同时缺乏成熟的 ADAM3 从附睾尾部精子中丢失。然而，PRSS37 如何调节 ADAM3 的成熟仍很大程度上不清楚。在这里，我们通过 GFP 免疫沉淀结合质谱法在 PRSS37-EGFP 敲入小鼠中确定了 PRSS37 相互作用组。三种分子伴侣（CLGN、CALR3 和 PDILT）和三种 ADAM 蛋白（ADAM2、ADAM6B 和 ADAM4）被鉴定与 PRSS37 相互作用。巧合的是，其中五个人（除了 ADAM4）已经被报道与 ADAM3 前体相互作用并调节其成熟。我们进一步证明 PRSS37 还与 ADAM3 前体直接相互作用，其缺乏会阻碍 PDILT 和 ADAM3 之间的结合。这可能导致 ADAM3 向精细胞表面的不正确易位，导致 PRSS37 缺失的成熟精子中 ADAM3 丢失。对关键精子质膜蛋白成熟机制的理解将为避孕和治疗不明原因男性不育症的新策略铺平道路。
