Evidence that aspartic proteinase is involved in the proteolytic processing event of procathepsin L in lysosomes

Nishimura, Yukio; Kawabata, Takahiro; Furuno, K.; Kato, Keitaro

doi:10.1016/0003-9861(89)90289-0

Cited by 68 publications

(27 citation statements)

References 34 publications

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“…2a) that corresponds to the intralysosomal processed form of cathepsin L (37). After treatment with PAN for 72 h, we found a strong up-regulation of cathepsin L and the induction of the 45-kDa non-processed pro-enzyme procathepsin L (38). Equal protein load was confirmed by reprobing the membrane with anti-␣-tubulin (Fig.…”

Section: Resultsmentioning

confidence: 65%

Podocyte Migration during Nephrotic Syndrome Requires a Coordinated Interplay between Cathepsin L and α3 Integrin

Reiser¹,

Oh²,

Shirato³

et al. 2004

Journal of Biological Chemistry

163

135

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Podocyte foot process effacement and disruption of the slit diaphragm are typically associated with glomerular proteinuria and can be induced in rats by the injection of puromycin aminonucleoside. Here, we show that the induction of puromycin aminonucleoside nephrosis involves podocyte migration conducted by a coordinated interplay between the cysteine protease cathepsin L and ␣ 3 integrin. Puromycin aminonucleoside treatment up-regulates cathepsin L expression in podocytes in vivo as well as expression and enzymatic activity of cathepsin L in podocytes in vitro. Isolated podocytes from mice lacking cathepsin L are protected from cell puromycin aminonucleoside-induced cell detachment. The functional significance of cathepsin L expression was underscored by the observation that puromycin aminonucleoside-induced cell migration was slowed down in cathepsin L-deficient podocytes and by the preservation of cell-cell contacts and expression of vital slit diaphragm protein CD2AP. Cathepsin L expression and activity were induced in podocytes lacking ␣ 3 integrin. Similarly, acute functional inhibition of ␣ 3 integrin in wild type podocytes with a blocking antibody increased the expression of cathepsin L activity. Downregulation of ␣ 3 integrin protected against puromycin aminonucleoside-induced podocyte detachment. In summary, these data establish that podocyte foot process effacement is a migratory event involving a novel interplay between cathepsin L and ␣ 3 integrin.Glomerular podocytes serve as a final barrier to urinary protein loss by the formation and maintenance of podocyte foot processes and the interposed slit diaphragm (SD) 1 All forms of nephrotic syndrome are characterized by podocyte foot process (FP) effacement and/or molecular reorganization of the slit diaphragm (1). FP effacement requires a precise interplay of multiple cellular functions including structural alterations of the cytoskeleton, movement of FP over the basement membrane, and reconstruction of the slit diaphragm (1). The discovery of several novel podocyte proteins and their mutation analysis including nephrin (2), CD2AP (3), ␣-actinin-4 (4), podocin (5), neph1 (6), and FAT (7) have shed light on the pathogenesis of FP effacement and proteinuria and emphasized the critical role of podocyte FP and the SD in maintaining the function of the glomerular filtration barrier.

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Section: Resultsmentioning

confidence: 65%

Podocyte Migration during Nephrotic Syndrome Requires a Coordinated Interplay between Cathepsin L and α3 Integrin

Reiser¹,

Oh²,

Shirato³

et al. 2004

Journal of Biological Chemistry

163

135

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“…Cathepsin L has multiple splice variants (34,36,37) and at least two isoforms (38,39). Like other acid hydrolases, cathepsin L is a zymogen that needs to be cleaved three times for full protease activity (35,(40)(41)(42)(43). Mock-or rgRSV-infected cells were harvested at 24 hpi in the absence of protease inhibitors and assayed for cathepsin L activity.…”

Section: Resultsmentioning

confidence: 99%

Preventing Cleavage of the Respiratory Syncytial Virus Attachment Protein in Vero Cells Rescues the Infectivity of Progeny Virus for Primary Human Airway Cultures

Corry

Johnson

Cornwell

et al. 2016

J Virol

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All live attenuated respiratory syncytial virus (RSV) vaccines that have advanced to clinical trials have been produced in Vero cells. The attachment (G) glycoprotein in virions produced in these cells is smaller than that produced in other immortalized cells due to cleavage. These virions are 5-fold less infectious for primary well-differentiated human airway epithelial (HAE) cell cultures. Because HAE cells are isolated directly from human airways, Vero cell-grown vaccine virus would very likely be similarly inefficient at initiating infection of the nasal epithelium following vaccination, and therefore, a larger inoculum would be required for effective vaccination. We hypothesized that Vero cell-derived virus containing an intact G protein would be more infectious for HAE cell cultures. Using protease inhibitors with increasing specificity, we identified cathepsin L to be the protease responsible for cleavage. Our evidence suggests that cleavage occurs in the late endosome or lysosome during endocytic recycling. Cathepsin L activity was 100-fold greater in Vero cells than in HeLa cells. In addition, cathepsin L was able to cleave the G protein in Vero cell-grown virions but not in HeLa cell-grown virions, suggesting a difference in G-protein posttranslational modification in the two cell lines. We identified by mutagenesis amino acids important for cleavage, and these amino acids included a likely cathepsin L cleavage site. Virus containing a modified, noncleavable G protein produced in Vero cells was 5-fold more infectious for HAE cells in culture, confirming our hypothesis and indicating the value of including such a mutation in future live attenuated RSV vaccines. IMPORTANCEWorldwide, RSV is the second leading infectious cause of infant death, but no vaccine is available. Experimental live attenuated RSV vaccines are grown in Vero cells, but during production the virion attachment (G) glycoprotein is cleaved. Virions containing a cleaved G protein are less infectious for primary airway epithelial cells, the natural RSV target. In the study described here we identified the protease responsible, located the cleavage site, and demonstrated that cleavage likely occurs during endocytic recycling. Moreover, we showed that the infectivity of Vero cell-derived virus for primary airway epithelial cells is increased 5-fold if the virus contains a mutation in the G protein that prevents cleavage. The blocking of cleavage should improve RSV vaccine yield, consequently reducing production costs. Posttranslational cleavage of the fusion glycoprotein of many viruses plays an essential role in activation; however, cleavage of the RSV G protein is a novel example of a detrimental effect of cleavage on virus infectivity. In adults and teenagers, most respiratory syncytial virus (RSV) infections produce upper respiratory tract infection and symptoms. However, in infants, the immunocompromised, and the elderly, RSV has the potential to cause life-threatening lower respiratory tract infection (1-4). Worldwide, in 2010 ...

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“…General protein degradation and turnover Many (including ↓) [36] Activation and degradation of polypeptide hormones, FGF [37] chemokines, growth factors and their receptors Plasminogen [38] Prolactin [39,40] Endostatin [41] Osteocalcin [42] Thyroglobulin [43] Insulin [44] Glucagon [45] IL-1 [46] IGFBP [47] Androgen receptor [48] Parathyroid hormone [49] MIP-1 alpha [50] MIP-1 beta [50] SLC [50] Activation of enzymatic precursors Cathepsin B [51,52] Cathepsin L [51,53] Transglutaminase 1 [54] Brain antigen processing Amyloid beta A4 protein [55,56] Tau [57] Tubulin [58] Myelin basic protein [59] Mhtt [60] Apolipoprotein E [61] Alpha-synuclein [62] Degradation of cytoskeletal proteins Neurofilaments [63] Actin [64,65] Myosin [64,66] Tropomyosin [65] Monocyte-mediated fibrinolysis Fibrinogen [67,68] Fibrin [67,68] Regulation of apoptosis Bid [69] Thiredoxin-1 [70] Processing of enzyme activators and inhibitors Prosaposin …”

Section: Biological Function Target Protein Referencementioning

confidence: 99%

Cathepsin D—Many functions of one aspartic protease

Beneš

Větvička

Fusek

2008

Critical Reviews in Oncology/Hematology

541

474

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For years, it has been held that cathepsin D (CD) is involved in rather nonspecific protein degradation in a strongly acidic milieu of lysosomes. Studies with CD knock-out mice revealed that CD is not necessary for embryonal development but it is indispensable for postnatal tissue homeostasis. Mutation that abolishes CD enzymatic activity causes neuronal ceroid lipofuscinosis (NCL) characterized by severe neurodegeneration, developmental regression, visual loss and epilepsy in both animals and humans.In the last decade, however, an increasing number of studies demonstrated that enzymatic function of CD is not restricted solely to acidic milieu of lysosomes with important consequences in regulation of apoptosis. In addition to CD enzymatic activity, it has been shown that apoptosis is also regulated by catalytically inactive mutants of CD which suggests that CD interacts with other important molecules and influences cell signaling.Moreover, procathepsin D, secreted from cancer cells, acts as a mitogen on both cancer and stromal cells and stimulates their pro-invasive and pro-metastatic properties. Numerous studies found that pCD/CD level represents an independent prognostic factor in a variety of cancers and is therefore considered to be a potential target of anti-cancer therapy.Studies dealing with functions of cathepsin D are complicated by the fact that there are several simultaneous forms of CD in a cell -pCD, intermediate enzymatically active CD and mature heavy and light chain CD. It became evident that these forms may differently regulate the above mentioned processes.In this article, we review the possible functions of CD and its various forms in cells and organisms during physiological and pathological conditions.

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Evidence that aspartic proteinase is involved in the proteolytic processing event of procathepsin L in lysosomes

Cited by 68 publications

References 34 publications

Podocyte Migration during Nephrotic Syndrome Requires a Coordinated Interplay between Cathepsin L and α3 Integrin

Podocyte Migration during Nephrotic Syndrome Requires a Coordinated Interplay between Cathepsin L and α3 Integrin

Preventing Cleavage of the Respiratory Syncytial Virus Attachment Protein in Vero Cells Rescues the Infectivity of Progeny Virus for Primary Human Airway Cultures

Cathepsin D—Many functions of one aspartic protease

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