A Mutation of Furin Causes the Lack of Precursor-Processing Activity in Human Colon Carcinoma LoVo Cells

Takahashi, Shouji; Kasai, Kazuhiko; Hatsuzawa, Kiyotaka; Kitamura, Noriaki; Misumi, Yoshio; Ikehara, Yukio; Murakami, Kenji; Nakayama, Kazuhisa

doi:10.1006/bbrc.1993.2146

Cited by 123 publications

(128 citation statements)

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“…To confirm this finding, we performed western blot analysis using the LoVo cells, which express (P)RR without functional furin. 12,13 Using the N-(P)RR antibody, LoVo cell lysates showed FL-(P)RR at 39 kDa, and conditioned medium from the LoVo cells showed NTF-(P)RR at 29 kDa. Both bands were significantly suppressed by transfection with siRNA against (P)RR (Figure 3b).…”

Section: Resultsmentioning

confidence: 99%

The (pro)renin receptor is cleaved by ADAM19 in the Golgi leading to its secretion into extracellular space

Yoshikawa

Aizaki

Kusano³

et al. 2011

Hypertens Res

113

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The (pro)renin receptor ((P)RR), which is a recently discovered molecule of the renin-angiotensin system, plays an important role in the development of cardiovascular diseases. However, the molecular properties and the subcellular distribution of (P)RR remain controversial. In this study, (P)RR-Venus in Chinese hamster ovary (CHO) cells ((P)RR-Venus-CHO) or endogenous (P)RR in human vascular smooth muscle cells (VSMC) were constitutively cleaved without any stimulation, and secretion of the aminoterminal fragment (NTF-(P)RR) into the media was determined using western blot analysis. Immunofluorescent analysis showed robust expression of (P)RR in the endoplasmic reticulum (ER) or the Golgi but not in the plasma membrane. Moreover, we identified ADAM19, which is expressed in the Golgi, as one of cleaving proteases of (P)RR. Transfected ADAM19 evoked the shedding of (P)RR, whereas transfected dominant negative ADAM19 suppressed it. Although (P)RR contains a furin cleavage site, neither the furin-deficient LoVo cells nor furin inhibitor-treated VSMC lost NTF-(P)RR in the media. The secreted NTF-(P)RR induced the renin activity of prorenin in the extracellular space. We describe that (P)RR is mainly localized in the subcellular organelles, such as the ER and Golgi, and (P)RR is cleaved by ADAM19 in the Golgi resulting in two fragments, NTF-(P)RR and CTF-(P)RR. These results may suggest that (P)RR is predominantly secreted into the extracellular space. Hypertension Research (2011) 34, 599-605; doi:10.1038/hr.2010.284; published online 27 January 2011Keywords: ADAM19; ectodomain shedding; (pro)renin receptor; renin-angiotensin system INTRODUCTION The (pro)renin receptor ((P)RR) is a recently discovered molecule of the renin-angiotensin system (RAS), which plays pivotal roles in the regulation of the cardiovascular system under normal and pathological conditions. (P)RR binds both renin and prorenin, which is the precursor form of renin. 1 Although the binding of renin to (P)RR may increase its catalytic activity, the binding affinity between (P)RR and renin is lower than that of (P)RR and prorenin. These results indicate that it may be necessary to focus on the interaction between (P)RR and prorenin. Prorenin does not display protease activity in the plasma because the enzymatic cleft is covered by the prosegment. 2 However, the binding of prorenin to (P)RR evokes the renin activity without removal of its prosegment. This nonproteolytic activation of prorenin contributes to the activation of the local RAS. In addition to the enzymatic activity, renin/prorenin has been shown to provide other (P)RR-mediated effects. 3 The binding of renin/prorenin to (P)RR induces the activation of intracellular signaling, including the p38 MAP kinase-HSP27 cascade, the PI3K pathway and the ERK 1/2 pathway; these effects occur independently of angiotensin II, which is a final product of RAS. 2,4,5 Although this evidence strongly supports (P)RR localization in the plasma membrane, the subcellular localization of (P)RR remains unknown. (P)R...

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Section: Resultsmentioning

confidence: 99%

The (pro)renin receptor is cleaved by ADAM19 in the Golgi leading to its secretion into extracellular space

Yoshikawa

Aizaki

Kusano³

et al. 2011

Hypertens Res

113

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“…In the presence of ␣ 1 -PDX, full-length 50-kDa EDA-HA protein was detected, whereas little or no 36͞27-kDa C-terminal fragments were found in either the cell layer or the medium. Second, EDA-HA was expressed in the colon carcinoma cell line LoVo, in which both alleles of the furin gene are mutated and the furin protein is nonfunctional (36). Only the full-length EDA-HA molecule was detected in the cell layer, and no 36͞27-kDa fragments were detected in the medium.…”

Section: Resultsmentioning

confidence: 99%

Mutations within a furin consensus sequence block proteolytic release of ectodysplasin-A and cause X-linked hypohidrotic ectodermal dysplasia

Chen

Molloy

Thomas

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

112

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“…The very recent discovery of PC7 [11] precluded its analysis in this report. For this purpose, we coexpressed each candidate convertase with the precursor substrates in either the kidney epithelial cell line BSC40 or the furin activity-deficient colon carcinoma cell line LoVo [19]. Pulse and/or pulse-chase data elucidated the biosynthetic pathway of proNT3 and proBDNF and their processing into NT3 and BDNF.…”

Section: Introductionmentioning

confidence: 99%

Cellular processing of the neurotrophin precursors of NT3 and BDNF by the mammalian proprotein convertases

et al. 1996

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In order to define the enzymes responsible for the maturation of the precursors of brain-derived neurotrophic factor (proBDNF) and neurotrophin-3 (proNT3), we have analysed their biosynthesis and intracellular processing by the proprotein convertases furin, PC1, PC2, PACE4, PC5 and its isoform PC5/6-B. In these studies, we utilized a vaccinia virus expression system in either BSC40 or the furin activity-deficient LoVo cells. Results demonstrated that in both cells furin and, to a lesser extent, PACE4 and PC516-B effectively process proBDNF and proNT3. Furthermore, we have determined that human proNT3 is sulfated, suggesting that processing of proNT3 occurs following the arrival of the precursor to the Trans Golgi Network.

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A Mutation of Furin Causes the Lack of Precursor-Processing Activity in Human Colon Carcinoma LoVo Cells

Cited by 123 publications

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The (pro)renin receptor is cleaved by ADAM19 in the Golgi leading to its secretion into extracellular space

The (pro)renin receptor is cleaved by ADAM19 in the Golgi leading to its secretion into extracellular space

Mutations within a furin consensus sequence block proteolytic release of ectodysplasin-A and cause X-linked hypohidrotic ectodermal dysplasia

Cellular processing of the neurotrophin precursors of NT3 and BDNF by the mammalian proprotein convertases

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