Zhang H, Wada J, Hida K, Tsuchiyama Y, Hiragushi K, Shikata K, Wang H, Lin S, Kanwar Y S, Makino H
Department of Medicine III, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 780-8558, Japan.
J Biol Chem. 2001 May 18;276(20):17132-9. doi: 10.1074/jbc.M006723200. Epub 2001 Jan 31.
Collectrin, a novel homolog of angiotensin-converting enzyme-related carboxypeptidase (ACE2), was identified during polymerase chain reaction-based cDNA subtraction and up-regulated in 5/6 ablated kidneys at hypertrophic phase. Collectrin, with 222 amino acids, has an apparent signal peptide and a transmembrane domain; the sequence is conserved in mouse, rat, and human and shares 81.9% identity. Human collectrin has 47.8% identity with non-catalytic extracellular, transmembrane, and cytosolic domains of ACE2; however, unlike ACE and ACE2, collectrin lacks active dipeptidyl carboxypeptidase catalytic domains. The collectrin mRNA transcripts are expressed exclusively in the kidney. In situ hybridization reveals its mRNA expression in renal collecting ducts, and immunohistochemistry shows that it is localized to the luminal surface and cytoplasm of collecting ducts. Immunoprecipitation studies, using [35S]methionine-labeled renal cortical and inner medullar collecting duct cells, i.e. M-1 and mIMCD-3, indicate that the protein size is approximately 32 kDa. During the development of mouse kidney, mRNA signal is detectable at day 13 of gestation, and the protein product is observed in the ureteric bud branches. Its expression is progressively increased during later stages of the gestation extending into the neonatal periods and then is decreased in adult life. Up-regulated expression of collectrin in the hypertrophic kidneys after renal ablation and restricted spatio-temporal expression during development indicates a possible role(s)in the process of progressive renal failure and renal organogenesis.
Collectrin是血管紧张素转换酶相关羧肽酶(ACE2)的一种新型同源物,在基于聚合酶链反应的cDNA消减过程中被鉴定出来,在5/6肾切除术后肥大期的肾脏中表达上调。Collectrin含有222个氨基酸,有一个明显的信号肽和一个跨膜结构域;该序列在小鼠、大鼠和人类中保守,同源性为81.9%。人类Collectrin与ACE2的非催化性细胞外、跨膜和胞质结构域有47.8%的同源性;然而,与ACE和ACE2不同的是,Collectrin缺乏活性二肽基羧肽酶催化结构域。Collectrin mRNA转录本仅在肾脏中表达。原位杂交显示其mRNA在肾集合管中表达,免疫组织化学表明它定位于集合管的管腔表面和细胞质。使用[35S]甲硫氨酸标记的肾皮质和内髓集合管细胞(即M-1和mIMCD-3)进行免疫沉淀研究表明,该蛋白大小约为32 kDa。在小鼠肾脏发育过程中,妊娠第13天可检测到mRNA信号,在输尿管芽分支中观察到蛋白产物。其表达在妊娠后期直至新生儿期逐渐增加,然后在成年期下降。肾切除术后肥大肾脏中Collectrin表达上调以及发育过程中时空表达受限表明其在进行性肾衰竭和肾器官发生过程中可能发挥作用。