A Second Mutant Allele of Furin in the Processing-incompetent Cell Line, LoVo

Takahashi, Shouji; Nakagawa, Tsutomu; Kasai, Kazuhiko; Banno, Tomohiro; Duguay, Stephen J.; Ven, Wim J.M. Van de; Murakami, Kazuo; Nakayama, Kazuhisa

doi:10.1074/jbc.270.44.26565

Cited by 122 publications

(104 citation statements)

References 46 publications

(41 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Furin-deficient human colon carcinoma LoVo cells (32) were used as a source of pro-␣ v . Pro-␣ v integrins were partially purified from LoVo cells on a mAb L230 column.…”

Section: Methodsmentioning

confidence: 99%

An Alternative Processing of Integrin αv Subunit in Tumor Cells by Membrane Type-1 Matrix Metalloproteinase

Ratnikov¹,

Rozanov²,

Postnova³

et al. 2002

Journal of Biological Chemistry

141

102

View full text Add to dashboard Cite

“…Furin-deficient human colon carcinoma LoVo cells (32) were used as a source of pro-␣ v . Pro-␣ v integrins were partially purified from LoVo cells on a mAb L230 column.…”

Section: Methodsmentioning

confidence: 99%

An Alternative Processing of Integrin αv Subunit in Tumor Cells by Membrane Type-1 Matrix Metalloproteinase

Ratnikov¹,

Rozanov²,

Postnova³

et al. 2002

Journal of Biological Chemistry

141

102

View full text Add to dashboard Cite

“…To examine the importance of furin in the processing of pro-CNP, we performed experiments using furin-deficient LoVo cells that were derived from a lymph node metastasis of a human colon adenocarcinoma and contain compound mutations in the furin gene (36). As shown in Fig.…”

Section: Effects Of Small Molecule Inhibitors On Processing Of Pro-cnp-mentioning

confidence: 99%

Furin-mediated Processing of Pro-C-type Natriuretic Peptide

Pan

et al. 2003

Journal of Biological Chemistry

187

110

View full text Add to dashboard Cite

C-type natriuretic peptide (CNP) is a member of the natriuretic peptide family that is involved in a variety of homeostatic processes. Here we characterize the processing essential for the conversion of the precursor, human pro-CNP, to the biologically active hormone. In human embryonic kidney 293 and chondrosarcoma SW 1353 cells, recombinant pro-CNP was converted into a mature peptide intracellularly as detected by Western analysis. Expression of recombinant human corin, a proatrial natriuretic peptide convertase, did not enhance the processing of pro-CNP in these cells. The processing of pro-CNP was inhibited in the presence of an inhibitor of the endoprotease furin but was not affected by inhibitors of matrix metalloproteinases and tumor necrosis factor-␣ convertase. In furin-deficient human colon adenocarcinoma LoVo cells, no conversion of recombinant pro-CNP to CNP was detected. Expression of recombinant human furin in LoVo cells restored the ability of these cells to process pro-CNP. Furthermore, incubation of purified recombinant human furin with LoVo cell lysate containing pro-CNP led to the conversion of the precursor to a mature peptide. The furin-processed CNP was shown to be biologically active in a cell-based cGMP assay. These results demonstrate that furin is a critical enzyme for the processing of human pro-CNP.The natriuretic peptide family consists of three structurally related peptides: atrial natriuretic peptide (ANP), 1 brain or B-type natriuretic peptide (BNP), and C-type natriuretic peptide (CNP) (1-7). ANP and BNP are produced mainly in cardiomyocytes in the heart and are important in maintaining normal body fluid and sodium homeostasis. CNP is expressed in many tissues and cell types, including the brain, vascular endothelial cells, and chondrocytes (8 -13). The dominant receptor for CNP is natriuretic peptide receptor-B, whereas the receptor for ANP and BNP is natriuretic peptide receptor-A. The biological functions of CNP are apparently different from those of ANP and BNP. Studies have shown that CNP inhibits the proliferation of vascular smooth muscle cells in culture (14, 15) and prevents balloon injury-induced coronary artery restenosis in animal models (16 -18). Recent studies of CNPdeficient mice indicate that CNP plays an important role in chondrocyte differentiation and bone formation (11).The natriuretic peptides are synthesized as prepropeptides. The signal peptide is removed to form propeptides, but a further proteolytic cleavage of the propeptide is required to convert the precursor to a biologically active peptide. This activation mechanism is critical in regulation of the activity of the natriuretic peptides, but the enzyme(s) responsible for the conversion remained uncharacterized for many years. Recently, we identified a cardiac serine protease, corin (19), that is a member of the type II transmembrane serine protease family (20,21). In cell-based functional studies, we showed that corin converted pro-ANP to biologically active ANP in a highly sequence-specific manner (22...

show abstract

“…In vitro casein kinase II (CK2) assays were performed according to the methods reported by Takahashi et al (20). Briefly, bacterially expressed GST fusion proteins were pre-bound to glutathione-Sepharose 4B beads (Amersham Biosciences).…”

Section: Fig 1 Phosphorylation Of Pc-2 In Vivomentioning

confidence: 99%

Calcium Dependence of Polycystin-2 Channel Activity Is Modulated by Phosphorylation at Ser812

Cai

Anyatonwu

Okuhara

et al. 2004

Journal of Biological Chemistry

139

188

View full text Add to dashboard Cite

Polycystin-2 (PC-2) is a non-selective cation channel that, when mutated, results in autosomal dominant polycystic kidney disease. In an effort to understand the regulation of this channel, we investigated the role of protein phosphorylation in PC-2 function. We demonstrated the direct incorporation of phosphate into PC-2 in cells and tissues and found that this constitutive phosphorylation occurs at Ser 812 , a putative casein kinase II (CK2) substrate domain. Ser 812 can be phosphorylated by CK2 in vitro and substitution S812A results in failure to incorporate phosphate in cultured epithelial cells. Non-phosphorylated forms of PC-2 traffic normally in the endoplasmic reticulum and cilial compartments and retain homo-and hetero-multimerization interactions with PC-2 and polycystin-1, respectively. Single-channel studies of PC-2, S812A, and a substitution mutant, T721A, not related to phosphorylation show that PC-2 and S812A function as divalent cation channels with similar current amplitudes across a range of holding potentials; the T721A channel is not functional. Channel open probabilities for PC-2 and S812A show a bell-shaped dependence on cytoplasmic Ca 2؉ but there is a shift in this Ca 2؉ dependence such that S812A is 10-fold less sensitive to Ca 2؉ activation/inactivation than the wild type PC-2 channel. In vivo analysis of PC-2-dependent enhanced intracellular Ca 2؉ transients found that S812A resulted in enhanced transient duration and relative amplitude intermediate between control cells and those overexpressing wild type PC-2. Phosphorylation at Ser 812 modulates PC-2 channel activity and factors regulating this phosphorylation are likely to play a role in the pathogenesis of polycystic kidney disease.

show abstract

A Second Mutant Allele of Furin in the Processing-incompetent Cell Line, LoVo

Cited by 122 publications

References 46 publications

An Alternative Processing of Integrin αv Subunit in Tumor Cells by Membrane Type-1 Matrix Metalloproteinase

An Alternative Processing of Integrin αv Subunit in Tumor Cells by Membrane Type-1 Matrix Metalloproteinase

Furin-mediated Processing of Pro-C-type Natriuretic Peptide

Calcium Dependence of Polycystin-2 Channel Activity Is Modulated by Phosphorylation at Ser812

Contact Info

Product

Resources

About