A furin-defective cell line is able to process correctly the gp160 of human immunodeficiency virus type 1

Ohnishi, Yasuharu; Shioda, Tatsuo; Nakayama, Kazuhisa; Iwata, Seiko; Gotoh, Bin; Hamaguchi, Michinari; Nagai, Yoshinori

doi:10.1128/jvi.68.6.4075-4079.1994

Cited by 80 publications

(47 citation statements)

References 41 publications

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“…Three rabbit antibodies were raised against BACE using peptides derived from the proregion (residues [26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45], the Nterminal end of the mature protein (residues 46-65), and the C-terminal 15 residues. The peptides BACE [26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45] C-IRLPLRSGLGGAPLGLRLPR, BACE 46-65 ETDEEPEEPGRRGSFVEMVD-C, and BACE CT15 C-RQQHDDFADDISLLK were synthesized at the Mayo Clinic protein core facility with terminal cysteine residues as indicated and coupled to maleimide-activated keyhole limpet hemocyanin (Pierce, Rockford, IL). Antibodies were raised at Cocalico Biologicals (Reamstown, PA) using a custom schedule, and the sera provided were tested by Western blot analysis as previously described (46).…”

Section: Antibodiesmentioning

confidence: 99%

“…Antibodies were raised at Cocalico Biologicals (Reamstown, PA) using a custom schedule, and the sera provided were tested by Western blot analysis as previously described (46). Antibodies against BACE [26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45] will not detect mature BACE, because the peptide is derived from the proregion of BACE that is lost upon proteolytic maturation ( Fig. 1).…”

Section: Antibodiesmentioning

confidence: 99%

“…Cells were lysed in MPER (Pierce), adjusted to RIPA (47), and immunoprecipitated using protein-A beads and a non-immune antiserum to reduce background. The supernatant was divided into two parts, precipitated with BACE [26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44][45] and BACE CT15 in RIPA buffer overnight, and separated on 4-15% polyacrylamide gels (Biorad, Richmond, CA). Western blot analyses of media and lysates were conducted and quantified as described previously using precast 10% NuPAGE (Invitrogen) or Biorad 4-15% gels (46).…”

Section: Immunoassaysmentioning

confidence: 99%

“…Although all these enzymes share a similar cleavage-sequence motif, they have distinct substrate preferences. For example, LoVo cells that are deficient in furin are incapable of cleaving Newcastle disease virus fusion glycoprotein but cleave Human Immunodeficiency Virus envelope protein, GP160, as efficiently as normal cell lines (39).…”

mentioning

confidence: 99%

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Convertases other than furin cleave β‐secretase to its mature form

Pinnix¹,

Roseberry²,

Onstead³

et al. 2001

FASEB j.

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An aspartyl protease, Beta‐Site APP cleaving enzyme (BACE), was identified as the β‐secretase responsible for generating the Amyloid β protein that is believed to cause Alzheimer's disease. BACE has a short propeptide domain that is absent in the mature enzyme because of proteolytic cleavage after the sequence RLPR. This sequence is a predicted substrate for proprotein convertases such as furin. To determine the role of furin and other proprotein convertases, we expressed proBACE in a furin‐deficient mutant Chinese hamster ovary (CHO‐K1) line, RPE.40. ProBACE signal was higher in RPE.40 than in the CHO‐K1 parent, which confirmed that furin plays a role in propeptide removal. However, two independent approaches showed that proBACE is cleaved to mature BACE in RPE.40: proBACE was rapidly turned over in RPE.40 although total BACE was stable, and decanoyl‐RVKR‐chloromethylketone, an inhibitor of the proprotein convertase family, substantially increased proBACE levels in both RPE40 and CHO‐K1. Transient transfection shows that furin, PACE4, PC5/6, and PC7 mediate BACE cleavage in vivo, at least when overexpressed. RPE.40 is proficient in BACE activity despite its furin deficiency. Therefore, our finding that proBACE is cleaved in this mutant leaves open the possibility that maturation is an important regulatory step and a therapeutic target.

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Section: Antibodiesmentioning

confidence: 99%

Section: Antibodiesmentioning

confidence: 99%

Section: Immunoassaysmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Convertases other than furin cleave β‐secretase to its mature form

Pinnix¹,

Roseberry²,

Onstead³

et al. 2001

FASEB j.

View full text Add to dashboard Cite

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“…Generation of a recombinant vaccinia virus expressing the HCV core (VV-core) was described [14]. Recombinant vaccinia viruses expressing the HCV C-E1 (VV-C-E1), E2 (VV-E2), NS3 (VV-NS3), NS4 (VV-NS4) or NS5 (VV-NS5) were generated as described [15]. In brief, the HCV C-E1, E2, NS3, NS4 and NS5 genes corresponding to amino acid residues 1-380, 381-810, 1027-1657, 1658-1972 and 1959-2872, respectively, were amplified from the plasmid pBRTM/HCV1-3011con which contains the entire DNA sequence derived from the HCV H77 clone [16] (kindly provided by Dr. Charles M. Rice, The Rockefeller University, New York, NY) by PCR and inserted into the transfer vector, pNZ68K2.…”

Section: Virusesmentioning

confidence: 99%

Coupling to the surface of liposomes alters the immunogenicity of hepatitis C virus-derived peptides and confers sterile immunity

Takagi

Kobayashi

Taneichi

et al. 2013

Biochemical and Biophysical Research Communications

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We have previously demonstrated that antigens chemically coupled to the surface of liposomes consisting of unsaturated fatty acids were cross-presented by antigen presenting cells to cytotoxic T lymphocytes (CTLs). Liposomal form of immunodominant CTL epitope peptides derived from lymphocytic choriomeningitis virus exhibited highly efficient antiviral CTL responses in immunized mice. In this study, we coupled 15 highly conserved immunodominant CTL epitope peptides derived from hepatitis C virus (HCV) to the surface of liposomes. We also emulsified the peptides in incomplete Freund's adjuvant, and compared the immune responses of the two methods of presenting the peptides by cytotoxicity induction and interferon-gamma (IFN-γ) production by CD8(+) T cells of the immunized mice. We noticed significant variations of the immunogenicity of each peptide between the two antigen delivery systems. In addition, the immunogenicity profiles of the peptides were also different from those observed in the mice infected with recombinant adenoviruses expressing HCV proteins as previously reported. Induction of anti-viral immunity by liposomal peptides was tested by the challenge experiments using recombinant vaccinia viruses expressing corresponding HCV epitopes. One D(b)-restricted and three HLA-A(*)0201-restricted HCV CTL epitope peptides on the surface of liposomes were found to confer complete protection to immunized mice with establishment of long-term memory. Interestingly, their protective efficacy seemed to correlate with the induction of IFN-γ producing cells rather than the cytotoxicity induction suggesting that the immunized mice were protected through non-cytolytic mechanisms. Thus, these liposomal peptides might be useful as HCV vaccines not only for prevention but also for therapeutic use.

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Furin‐Mediated Sequential Delivery of Anticancer Cytokine and Small‐Molecule Drug Shuttled by Graphene

et al. 2014

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Graphical abstract A cellular protease-mediated graphene-based nanosystem is developed for co-delivery of a membrane-associated cytokine (TRAIL) and an intracellular-acting small-molecule drug (DOX). The nanocarrier realizes the intramembrane enzyme-mediated extracellular release of TRAIL and endocytotic acidity-responsive intracellular release of DOX, which enables them to target to their distinct sites of action. This formulation starts a new generation of 2D nanomaterials with programmed-release therapeutics capability for combination cancer treatment.

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A furin-defective cell line is able to process correctly the gp160 of human immunodeficiency virus type 1

Cited by 80 publications

References 41 publications

Convertases other than furin cleave β‐secretase to its mature form

Convertases other than furin cleave β‐secretase to its mature form

Coupling to the surface of liposomes alters the immunogenicity of hepatitis C virus-derived peptides and confers sterile immunity

Furin‐Mediated Sequential Delivery of Anticancer Cytokine and Small‐Molecule Drug Shuttled by Graphene

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