Biosynthesis and Post-translational Processing of the Precursor to Brain-derived Neurotrophic Factor

Pourquoi ce travail est-il dans la base ?

Une base qui oublie comment elle a trouvé un travail ne peut pas être vérifiée. Voici les voies qui ont admis celui-ci.

Aucune affiliation canadienne. Une base fondée sur la seule affiliation (le devis habituel) n'aurait jamais vu ce travail. C'est l'un des travaux qui justifient l'inversion de la base.

Scores machine (provisoires)

Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.

Écart entre enseignants

0,230 · la distance entre les deux têtes enseignantes sur ce seul travail

Statut de validation

score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découle

Résumé

We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. Some pro-BDNF is released extracellularly and is biologically active as demonstrated by its ability to mediate TrkB phosphorylation. The precursor undergoes N-terminal cleavage within the trans-Golgi network and/or immature secretory vesicles to generate mature BDNF (14 kDa). Small amounts of a 28-kDa protein that is immunoprecipitated with BDNF antibodies is also evident. This protein is generated in the endoplasmic reticulum through N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr57-↓-Ser-Leu site. Cleavage is abolished when Arg54 is changed to Ala (R54A) by in vitromutagenesis. Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with α1-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. We examined the biosynthesis and post-translational processing of the brain-derived neurotrophic factor precursor (pro-BDNF) in cells infected with a pro-BDNF-encoding vaccinia virus. Metabolic labeling, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis reveal that pro-BDNF is generated as a 32-kDa precursor that is N-glycosylated and glycosulfated on a site, within the pro-domain. Some pro-BDNF is released extracellularly and is biologically active as demonstrated by its ability to mediate TrkB phosphorylation. The precursor undergoes N-terminal cleavage within the trans-Golgi network and/or immature secretory vesicles to generate mature BDNF (14 kDa). Small amounts of a 28-kDa protein that is immunoprecipitated with BDNF antibodies is also evident. This protein is generated in the endoplasmic reticulum through N-terminal cleavage of pro-BDNF at the Arg-Gly-Leu-Thr57-↓-Ser-Leu site. Cleavage is abolished when Arg54 is changed to Ala (R54A) by in vitromutagenesis. Blocking generation of 28-kDa BDNF has no effect on the level of mature BDNF and blocking generation of mature BDNF with α1-PDX, an inhibitor of furin-like enzymes, does not lead to accumulation of the 28-kDa form. These data suggest that 28-kDa pro-BDNF is not an obligatory intermediate in the formation of the 14-kDa form in the constitutive secretory pathway. Brain-derived neurotrophic factor (BDNF) 1The abbreviations used are:BDNFbrain-derived neurotrophic factorNGFnerve growth factorNT-3neurotrophin-3NT-4/5neurotrophin-4/5PAGEpolyacrylamide gel electrophoresisvvvaccinia virusendo Hendoglycosidase HERendoplasmic reticulumα1-PDXα1-antitrypsin PortlandDMEMDulbecco's modified Eagle's mediumrhBDNFrecombinant human BDNF along with nerve growth factor (NGF), neurotrophin-3 (NT-3), and neurotrophin-4/5 (NT-4/5) are members of the neurotrophin family of trophic factors (1Snider D.W. Cell. 1994; 77: 625-638Abstract Full Text PDF PubMed Scopus (1298) Google Scholar). The neurotrophins play essential roles in the development, survival, and function of a wide range of neurons in both the peripheral and central nervous systems. brain-derived neurotrophic factor nerve growth factor neurotrophin-3 neurotrophin-4/5 polyacrylamide gel electrophoresis vaccinia virus endoglycosidase H endoplasmic reticulum α1-antitrypsin Portland Dulbecco's modified Eagle's medium recombinant human BDNF The neurotrophins have a number of shared characteristics, including similar molecular weights (13.2–15.9 kDa), isoelectric points (in the range of 9–10), and ∼50% identity in primary structure. They exist in solution as noncovalently bound dimers. Six cysteine residues conserved in the same relative positions give rise to three intra-chain disulfide bonds (2Maisonpierre P.C. Belluscio L. Squito S. Ip N.Y. Furth M.E. Lindsay R.M. Yancoupoulos G.D. Science. 1990; 247: 1373-1520Crossref Scopus (1108) Google Scholar, 3Maisonpierre P.C. Le Beau M.M. Espinosa III, R. Ip N.Y. Belluscia L. De La Monte S.M. Squinto S. Furth M.E. Yancopoulos G.D. Genomics. 1991; 10: 558-568Crossref PubMed Scopus (441) Google Scholar). The neurotrophins interact with two cell surface receptors, the low affinity P75 receptor (4Barker P.A. Cell Death Differ. 1998; 5: 346-356Crossref PubMed Scopus (123) Google Scholar) and the Trk family of high affinity tyrosine kinase receptors (5Kaplan D.R. Hemstead B.L. Martin-Zanca D. Chao M.V. Parada L.F. Science. 1991; 252: 545-558Crossref Scopus (1132) Google Scholar). NGF preferentially binds TrkA, BDNF and NT4/5 bind TrkB, and NT-3 binds TrkC (and TrkA to a lesser extent). Sequence data predict that mature neurotrophins are generated through the proteolytic processing of higher molecular weight precursors (31–35 kDa), a process that has been extensively studied with respect to the production of NGF (6Edwards R.H. Selby M.J. Mobley W.C. Weinrich S.L. Hruby D.E. Rutter W.J. Mol. Cell. Biol. 1988; 8: 2456-2464Crossref PubMed Scopus (89) Google Scholar, 7Seidah N.G. Benjannet S. Pareek S. Savaria D. Hamlin J. Goulet B. Laliberte J. Lazure C. Chretien M. Murphy R.A. Biochem. J. 1996; 314: 951-960Crossref PubMed Scopus (239) Google Scholar). Almost nothing is known, however, about the biosynthesis and post-translational processing of the other members of the neurotrophin family. Recent data from our laboratory show that cells with a regulated secretory pathway, including central nervous system neurons, release mature (i.e. fully processed) NGF (8Mowla S.J. Pareek S. Farhadi H.F. Petrecca K. Fawcett J.P. Seidah N.G. Morris S.J. Sossin W.S. Murphy R.A. J. Neurosci. 1999; 19: 2069-2080Crossref PubMed Google Scholar) and NT-3 (9Farhadi H.F. Mowla S.J. Petrecca K. Morris S.J. Seidah N.G. Murphy R.A. J. Neurosci. 2000; 20: 4059-4068Crossref PubMed Google Scholar) via the constitutive secretory pathway, whereas mature BDNF is packaged in vesicles and released through the regulated pathway (8Mowla S.J. Pareek S. Farhadi H.F. Petrecca K. Fawcett J.P. Seidah N.G. Morris S.J. Sossin W.S. Murphy R.A. J. Neurosci. 1999; 19: 2069-2080Crossref PubMed Google Scholar). Furthermore, BDNF is contained in a microvesicular fraction of lysed brain synaptosomes consistent with its anterograde transport in large dense core vesicles (10Fawcett J.P. Aloyz R. McClean J.H. Pareek S. Miller F.D. McPherson P.S. Murphy R.A. J. Biol. Chem. 1997; 272: 8837-8840Abstract Full Text Full Text PDF PubMed Scopus (103) Google Scholar). Differences in the intracellular sorting of neurotrophins may arise, at least in part, from differences in the chemistry and processing of their precursors. Therefore, defining how neurotrophins are generated within a cell will be key to understanding how neurotrophins are released and function within the nervous system. In this study, we monitored the biosynthesis and post-translational processing of the precursor to BDNF (pro-BDNF) using a vaccinia virus (vv) expression system together with metabolic labeling, immunoprecipitation, and SDS-PAGE. Data show that pro-BDNF is produced as a 32-kDa precursor that undergoes N-glycosylation and glycosulfation on residues located within the pro-domain of the precursor. N-terminal cleavage of the precursor generates mature BDNF as well as a minor truncated form of the precursor (28 kDa) that arises by a different processing mechanism than mature BDNF. Site-directed mutagenesis data suggest that 28-kDa BDNF is not an obligatory intermediate in the formation of the mature form. Data also demonstrate that pro-BDNF could be biologically active, as determined by its ability to promote TrkB autophosphorylation. Hippocampal neurons were prepared according to the method of Brewer et al. (29Brewer G.J. Torricelli J.R. Evege E.K. Price P.J. J. Neurosci. Res. 1993; 35: 567-576Crossref PubMed Scopus (1883) Google Scholar). Briefly, the hippocampus was dissected from day 18 (E18) mice (Charles River Breeding Laboratories, Montreal, Canada), exposed to trypsin, dissociated mechanically, and grown in 60-mm collagen/poly-l-lysine-coated dishes. Cultures were maintained in serum-free Neurobasal medium (Life Technologies, Inc.) containing 0.5 mm glutamine and 1× supplemented B27 (Life Technologies, Inc.). AtT-20, COS-7, and LoVo cells were cultured as reported previously (7Seidah N.G. Benjannet S. Pareek S. Savaria D. Hamlin J. Goulet B. Laliberte J. Lazure C. Chretien M. Murphy R.A. Biochem. J. 1996; 314: 951-960Crossref PubMed Scopus (239) Google Scholar). A human glioma (U373) cell line and a variant (U373/PDX) that stably expresses α1-PDX, an inhibitor of furin-like enzymes (11Anderson E.D. Thomas L. Hayflick J.S. Thomas G. J. Biol. Chem. 1993; 268: 24887-24891Abstract Full Text PDF PubMed Google Scholar), was generously provided by Dr. Gary Thomas (Vollum Institute, Portland OR). Purified recombinant vv containing the full-length coding region of human pro-BDNF was prepared and used to infect cells as described previously (12Seidah N.G. Benjannet S. Pareek S. Chretien M. Murphy R.A. FEBS Lett. 1996; 379: 248-250Crossref Scopus (279) Google Scholar). U373 and U373/PDX glial cells and AtT-20 cells were grown in 60-mm dishes and exposed to virus for 30 min or 2 h, respectively. The cells were incubated in medium without virus overnight and either pulsed or pulse-chase labeled at 37 °C for specified time intervals. For pulse-chase experiments, infected cells were incubated in cysteine/methionine-free Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal calf serum for 1 h, and then received 1.5 ml of the same medium containing 0.2 mCi/ml [35S]Cys/Met (PerkinElmer Life Sciences) for 30 min. For the chase, cells were bathed for specified intervals in DMEM containing 10% fetal calf serum plus excess (2 times) cysteine and methionine. In experiments assessing sulfation, AtT-20 cells were labeled for 3 h with [Na235SO4 ] (0.5 mCi) (PerkinElmer Life Sciences) in methionine/cysteine/SO4-free RPMI 1640 medium (Life Technologies, Inc.). Sodium chlorate (1 mm) was added to the medium in some experiments to inhibit sulfation, and in others, tunicamycin (5 μg/ml) was added to inhibit N-linked glycosylation. In both cases, the drugs were present in the medium during the 60-min preincubation period and throughout the pulse-chase period. Radiolabeled BDNF was immunoprecipitated from cell lysates and conditioned medium as described previously (8Mowla S.J. Pareek S. Farhadi H.F. Petrecca K. Fawcett J.P. Seidah N.G. Morris S.J. Sossin W.S. Murphy R.A. J. Neurosci. 1999; 19: 2069-2080Crossref PubMed Google Scholar). We used an affinity-purified antibody to BDNF (13, kindly supplied by Amgen) at a of 0.5 were by SDS-polyacrylamide gel electrophoresis were for 1 h in and 10% with (PerkinElmer Life Sciences) for 1 h, in 10% for 1 h, and for h at was on from conditioned medium that were and from was using an as described previously S. C. Chretien M. Seidah N.G. J. Biol. Chem. Full Text PDF PubMed Google Scholar). AtT-20 cells were labeled with conditioned medium was and with antibody to and the were in of with or without H or were incubated overnight at 37 H were in and in mm containing mm using the (Life Technologies, we cells with of either the or form of a in and the cells were incubated for h in DMEM plus 10% fetal calf cells were labeled for h, and cell lysates and conditioned were and by SDS-PAGE. For we of the BDNF precursor by LoVo an cell line that is in furin-like enzymes S. K. K. K. K. Biochem. Res. 1993; PubMed Scopus Google Scholar), with and (11Anderson E.D. Thomas L. Hayflick J.S. Thomas G. J. Biol. Chem. 1993; 268: 24887-24891Abstract Full Text PDF PubMed Google Scholar). The cells were incubated in medium for h, by a in serum-free was for fully BDNF generated similar we LoVo cells with and to that the precursor was and we conditioned medium h from and vaccinia LoVo cells were used as for we used cells that TrkB, prepared and generously provided by Dr. The cells were bathed in conditioned medium for cell lysates were immunoprecipitated with antibody B.L. S.J. L. S. Parada L.F. D.R. Full Text PDF PubMed Scopus Google Scholar). The were in by using an and a for The were overnight at °C with a antibody in supplemented with and for an 1 h with a antibody was using (PerkinElmer Life the of the BDNF antibody and to its in we BDNF in AtT-20 cells using a vv the full-length precursor to of cell and conditioned medium were and immunoprecipitated with antibody to or BDNF antibody with excess (5 in in both cell and conditioned the antibody to BDNF immunoprecipitated three at and of with and were immunoprecipitated in the of excess We that the 32-kDa protein is the 28-kDa protein a truncated form of and the 14-kDa protein fully mature BDNF. the of the different of the BDNF precursor to mature we pulse-chase using AtT-20 cells infected with recombinant a system we have used previously (8Mowla S.J. Pareek S. Farhadi H.F. Petrecca K. Fawcett J.P. Seidah N.G. Morris S.J. Sossin W.S. Murphy R.A. J. Neurosci. 1999; 19: 2069-2080Crossref PubMed Google Scholar, H.F. Mowla S.J. Petrecca K. Morris S.J. Seidah N.G. Murphy R.A. J. Neurosci. 2000; 20: 4059-4068Crossref PubMed Google Scholar). 2 that 32-kDa BDNF precursor is in cell lysates as as min the cells were and in through 30 min of In cells labeled for a higher molecular weight is that a in cells for This and of the BDNF precursor the the 32-kDa to a lesser the minor higher molecular weight in whereas of the 14-kDa mature BDNF a The 28-kDa as as min and its level by 1 h of The of the 2 that amounts of the 32-kDa BDNF the 28-kDa and mature BDNF are released conditioned medium during the period. of the 32-kDa BDNF precursor and the truncated 28-kDa form of the precursor their to and that has no effect on the molecular of mature BDNF (14 kDa). with high the and 28-kDa BDNF that the precursor released the conditioned medium a of and high that both pro-BDNF and 28-kDa BDNF as with the whereas the higher is the of in the generation of BDNF from its we infected AtT-20 cells with and labeled the cells in the or of of an inhibitor of were labeled for 30 min by a in the or of that tunicamycin the of the BDNF precursors as well as mature BDNF the level of in the and of In the molecular of the BDNF precursor in cell and in conditioned medium was from to The that may play an in the BDNF precursor during its processing and not the molecular of mature BDNF (14 kDa), as this form of the protein is not Metabolic of AtT-20 cells with that pro-BDNF as well as the truncated 28-kDa form of the precursor are in is not Furthermore, of the with the that on is essential for the processing and/or of we labeled AtT-20 cells pro-BDNF with for 30 min and then the cells for 2 h in the or of chlorate (1 mm) P.A. Biochem. Res. PubMed Scopus Google Scholar). This by the that were protein in conditioned medium at the of the period not The that to chlorate no effect on processing of pro-BDNF or on of mature BDNF not in the cell the 28-kDa form of BDNF is we labeled infected cells with for 3 h in the or of A a that anterograde transport from the M. B. Mol. Biol. Cell. 1997; 8: PubMed Scopus Google Scholar). The cells were or a period without that no effect on the generation of the 28-kDa form of inhibit the generation of the 14-kDa form of mature BDNF. This effect was when the cells were 2 h in the of These suggest that the 28-kDa form of BDNF be generated in the whereas the mature form of as (8Mowla S.J. Pareek S. Farhadi H.F. Petrecca K. Fawcett J.P. Seidah N.G. Morris S.J. Sossin W.S. Murphy R.A. J. Neurosci. 1999; 19: 2069-2080Crossref PubMed Google Scholar), is generated in the trans-Golgi network or a The data show that cell lysates and conditioned of AtT-20 cells infected with generate a truncated form of BDNF with an molecular of We also this in other cell as well as in primary of neurons infected with the same vv In a study, we that a is to the level of 28-kDa BDNF when with pro-BDNF in cells N.G. Mowla S.J. J. Benjannet S. J.S. J. M. Lazure C. Murphy R.A. Chretien M. M. S. 1999; PubMed Scopus Google Scholar). N-terminal of 28-kDa BDNF a cleavage in 28-kDa BDNF is also at the same site, we at the to This could as a for this of N.G. Chretien M. Res. 1999; PubMed Scopus Google Scholar). of the pro-BDNF in of mature 14-kDa BDNF with no generation of the 28-kDa protein This that the protein is at the same site, and as for other at the is for cleavage of the cleavage within pro-BDNF that generates 28-kDa BDNF. N-terminal of 28-kDa BDNF. The N-terminal of the 28-kDa in cells infected with and a at positions and This that 28-kDa BDNF is generated by a cleavage at in the expression of the and form of pro-BDNF in cells were with expression of the or the Arg54 to Ala (R54A) form of cells were labeled with for h, and cell lysates and conditioned were immunoprecipitated with a and by In this study, we a vv pro-BDNF a cell line glial that stably expresses the inhibitor (10Fawcett J.P. Aloyz R. McClean J.H. Pareek S. Miller F.D. McPherson P.S. Murphy R.A. J. Biol. Chem. 1997; 272: 8837-8840Abstract Full Text Full Text PDF PubMed Scopus (103) Google Scholar). that furin-like enzymes the formation of mature BDNF has no effect on the generation of the 28-kDa as in expression of the Arg54 Ala in cells the generation of 28-kDa BDNF without the level of mature BDNF. suggest that the 28-kDa does not an obligatory intermediate in the processing of the BDNF precursor in the constitutive secretory pathway. amounts of pro-BDNF are conditioned our a that to the released in could be biologically this we to generate pro-BDNF by LoVo are in S. K. K. K. K. Biochem. Res. 1993; PubMed Scopus Google Scholar), with along with blocking the of furin-like enzymes in the we were to conditioned medium containing pro-BDNF and the 28-kDa BDNF without amounts of mature BDNF a for this study, we medium from LoVo cells with and vv (7Seidah N.G. Benjannet S. Pareek S. Savaria D. Hamlin J. Goulet B. Laliberte J. Lazure C. Chretien M. Murphy R.A. Biochem. J. 1996; 314: 951-960Crossref PubMed Scopus (239) Google Scholar), that the processing of pro-BDNF to mature BDNF medium from cells contained amounts of that medium from both cell TrkB in cells that the TrkB conditioned by cells infected with vv no We from data that released from a the BDNF precursor containing amounts of the 28-kDa form of BDNF has the to be biologically We not the of the 28-kDa form of pro-BDNF to this we were to amounts of the protein for in the of pro-BDNF or mature BDNF. biosynthesis of neurotrophins at low in neurons and is to neurotrophin processing with Therefore, in this study, we used a vaccinia virus expression system to pro-BDNF and to its processing in a of cell as well as in primary of We have used similar previously to the biosynthesis and post-translational processing of (7Seidah N.G. Benjannet S. Pareek S. Savaria D. Hamlin J. Goulet B. Laliberte J. Lazure C. Chretien M. Murphy R.A. Biochem. J. 1996; 314: 951-960Crossref PubMed Scopus (239) Google Scholar). using the BDNF antibody provided by as well as a antibody from not we three in cells and neurons, and 14-kDa of the protein that pro-BDNF is as a 32-kDa precursor that is within 1 h to give rise to mature BDNF (14 kDa). We also a of pro-BDNF released conditioned medium by AtT-20 cells and neurons (8Mowla S.J. Pareek S. Farhadi H.F. Petrecca K. Fawcett J.P. Seidah N.G. Morris S.J. Sossin W.S. Murphy R.A. J. Neurosci. 1999; 19: 2069-2080Crossref PubMed Google Scholar), cells that release both by the regulated and constitutive secretory In we not precursor release when similar were used to processing and release of the precursors of NGF (8Mowla S.J. Pareek S. Farhadi H.F. Petrecca K. Fawcett J.P. Seidah N.G. Morris S.J. Sossin W.S. Murphy R.A. J. Neurosci. 1999; 19: 2069-2080Crossref PubMed Google Scholar) or NT-3 (9Farhadi H.F. Mowla S.J. Petrecca K. Morris S.J. Seidah N.G. Murphy R.A. J. Neurosci. 2000; 20: 4059-4068Crossref PubMed Google Scholar). by M. B. Mol. Biol. Cell. 1997; 8: PubMed Scopus Google Scholar, G. J. Biol. Chem. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar) has that large amounts of the precursors of released by the regulated secretory pathway, as and are also released from AtT-20 differences could of pro-BDNF the sorting in the trans-Golgi network (8Mowla S.J. Pareek S. Farhadi H.F. Petrecca K. Fawcett J.P. Seidah N.G. Morris S.J. Sossin W.S. Murphy R.A. J. Neurosci. 1999; 19: 2069-2080Crossref PubMed Google Scholar), constitutive release of the precursor could also be of In that BDNF is present in the of neurons in J. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, M. J. Neurosci. 1997; PubMed Google Scholar), and as we nothing about the chemistry or of the BDNF protein within is that pro-BDNF could be produced in and in part, in an for as In this study, we have that medium containing pro-BDNF with the TrkB receptor and its the of pro-BDNF by cell the cells were exposed to a medium containing a large excess of This the of at the cell surface and has been to be for the P.A. Biochem. Res. PubMed Scopus Google Scholar). R. S. J. Biochem. 1994; PubMed Scopus Google Scholar) has that a BDNF containing an of of the cleavage of mature BDNF is biologically and R.H. Selby M.J. Rutter W.J. J. Biol. Chem. 1988; Full Text PDF PubMed Google Scholar) have reported that is biologically active at a level that of mature suggest that processing of may not be an for through the secretory pathway, the BDNF precursor is at the for N-linked residues of the cleavage that generates mature BDNF. This is conserved in the same in a for N-linked in neurotrophin and/or is released conditioned medium as a of and the and 28-kDa of BDNF as the and the is The of in the of has been well R. S. Biochem. Scopus Google Scholar). In the of with tunicamycin the of the and its (7Seidah N.G. Benjannet S. Pareek S. Savaria D. Hamlin J. Goulet B. Laliberte J. Lazure C. Chretien M. Murphy R.A. Biochem. J. 1996; 314: 951-960Crossref PubMed Scopus (239) Google Scholar). In this study, blocking N-glycosylation of pro-BDNF the level of of both pro-BDNF and mature may be to the of also demonstrate that to the pro-domain of the BDNF precursor are as has been previously reported for (7Seidah N.G. Benjannet S. Pareek S. Savaria D. Hamlin J. Goulet B. Laliberte J. Lazure C. Chretien M. Murphy R.A. Biochem. J. 1996; 314: 951-960Crossref PubMed Scopus (239) Google Scholar). Blocking with chlorate P.A. Biochem. Res. PubMed Scopus Google Scholar) not processing and release of This is consistent with the of and G.J. Mol. Cell. 1997; PubMed Scopus Google Scholar) that protein is not for the or proteolytic processing of to the regulated secretory pathway. We have also a 28-kDa protein that is a cleavage of the BDNF precursor in to 14-kDa mature BDNF that is generated through a processing pathway. Furthermore, its processing in U373 glial cells is not by α1-PDX, an inhibitor of the furin-like enzymes that generate mature BDNF from pro-BDNF in cells that a constitutive not regulated secretory pathway. These suggest that the 28-kDa is not by the by some other processing system within the Recent from our N.G. Mowla S.J. J. Benjannet S. J.S. J. M. Lazure C. Murphy R.A. Chretien M. M. S. 1999; PubMed Scopus Google Scholar, S.J. M. Seidah N.G. Murphy R.A. Neurosci. 1999; Scholar) that the 32-kDa BDNF precursor is a for a of pro-BDNF and produced 28-kDa BDNF for N-terminal that cleavage at the 28-kDa BDNF we is generated at the same cleavage site, we at the relative to the cleavage and is for by this of N.G. Chretien M. Res. 1999; PubMed Scopus Google Scholar). of the pro-BDNF not amounts of 28-kDa that the 28-kDa precursor is at the same the 28-kDa form of pro-BDNF is in our including neurons, we not the protein is biologically of suggest that the generation of mature BDNF in the constitutive pathway does not processing of pro-BDNF to the 28-kDa form. in the cell line constitutive cell line generation of 14-kDa BDNF is is no accumulation of 28-kDa as be the were an intermediate Ala of the abolished the generation of the 28-kDa form without the production of mature BDNF is to be about the BDNF precursor. For we not the precursor kDa) and the 28-kDa form of the both of be released from could have roles of their from mature BDNF. Furthermore, in cells with both the regulated and constitutive secretory pro-BDNF is preferentially and released from the regulated pathway, whereas (8Mowla S.J. Pareek S. Farhadi H.F. Petrecca K. Fawcett J.P. Seidah N.G. Morris S.J. Sossin W.S. Murphy R.A. J. Neurosci. 1999; 19: 2069-2080Crossref PubMed Google Scholar) and (9Farhadi H.F. Mowla S.J. Petrecca K. Morris S.J. Seidah N.G. Murphy R.A. J. Neurosci. 2000; 20: 4059-4068Crossref PubMed Google Scholar) are in the constitutive secretory pathway. may well of differences in the of the neurotrophin precursors or of are We for the antibody brain-derived neurotrophic We Benjannet and Lazure for pro-BDNF

Récupéré en direct depuis OpenAlex et désinversé. Les résumés ne sont pas conservés dans cette base de données : les index inversés représentent 8,6 Go des 9,3 Go de texte de la base, et le serveur dispose de 13 Go libres.

La notice

Revue

Journal of Biological Chemistry

Thématique

Nerve injury and regeneration

Domaine

Neuroscience

Établissements canadiens

—

Organismes subventionnaires

—

Mots-clés

ImmunoprecipitationTropomyosin receptor kinase BEndoplasmic reticulumBrain-derived neurotrophic factorNeurotrophic factorsFurinMolecular biologyGel electrophoresisBiochemistryChemistryCell biologyPolyacrylamide gel electrophoresisBiologyGeneEnzymeReceptor

Résumé présent dans OpenAlex

oui

API

/api/v1/works/W2148950534