Wolfgang Garten - Academia.edu (original) (raw)
Papers by Wolfgang Garten
PubMed, Feb 9, 2000
Antibodies specific to the N-terminal 34-aminoacid peptide of the major nucleocapsid protein NP o... more Antibodies specific to the N-terminal 34-aminoacid peptide of the major nucleocapsid protein NP of human influenza A viruses were obtained. The NP proteolytic cleavage occurring in infected cells late in infection has been demonstrated using this antibody. The antibody specifically reacted with noncleaved NP of 56 kD m.w. but not with cleaved NP of 53 kD m.w. This indicates that NP56-->NP53 intracellular cleavage released the N-terminal 3 kD peptide of Np molecule. The released 3 kD peptide did not accumulate in infected cells. Since the RNA-binding and nucleus migrating signals are located in the N-terminus of NP molecule, presumably, the terminal cleavage is important for intracellular NP functions.
SUMMARY Processing of the hemagglutinin involves transport from the rough endoplasmic reticulum t... more SUMMARY Processing of the hemagglutinin involves transport from the rough endoplasmic reticulum to the plasma membrane, glycosylation, and proteolytic cleavage. The tight coupling of these events is demonstrated in experiments in which mutants of fowl plague virus (FPV) with a temperature sensitive defect in transport have been analyzed. Two groups of such mutants have been characterized. With the first group (ts1, ts227), the hemagglutinin is arrested in the rough endoplasmic reticulum. With the second group (ts482, 532, 651), the hemagglutinin migrates to the Golgi apparatus, but does not reach the plasma membrane. The distribution of the carbohydrate side chains on the FPV hemagglutinin has been elucidated. HA 1 has 4 type I side chains attached to asparagine residues 12, 28, 123, and 149. The potential attachment site at asparagine 231 is not glycosylated. HA2 has a type II chain at asparagine 406 and a type I chain at asparagine 478. Comparison with other hemagglutinins demonstrates that the side chains in positions 12, 28, and 478 are conserved. The cleavage site has been analyzed with H3, H7, and H10 hemagglutinins. The available evidence indicates that proteolytic activation involves the action of a trypsin-like protease followed by the action of an exopeptidase of the carboxypeptidase B type: After in vitro cleavage with trypsin, v^iich is paralleled by activation of infectivity, the N-terminus of HA 2 and presumably the C-terminus of HA. are identical to those obtained after in vivo cleavage. This indicates that the carboxypeptidase B activity is present in the virion. After cleavage with the non- activating enzymes thermolysin and chymo trypsin, the cleavage site is shifted by a few amino acids. These observations indicate that activation of infectivity requires a highly specific amino acid sequence at the cleavage site.
Cold Spring Harbor Monograph Archive, 1994
Endoproteolytic cleavage, usually at arginine residues, is a common posttranslational modificatio... more Endoproteolytic cleavage, usually at arginine residues, is a common posttranslational modification of membrane and secretory proteins on the exocytotic transport route. Such proteins include precursors of peptide hormones, neuropeptides, growth factors, coagulation factors, serum albumin, cell-surface receptors, and adhesion molecules. All of these proteins play important roles in a large variety of different biological processes, and these functions depend on proteolytic cleavage of the proteins (for review, see Barr 1991). The same type of processing has also been observed with many viral membrane proteins (Fig. 1) and, in some instances, proved to be a crucial factor in determining organ and host tropism, spread of infection, and pathogenicity of these viruses. VIRUSES WITH SPIKE PROTEINS UNDERGOING POSTTRANSLATIONAL PROTEOLYTIC CLEAVAGE Paramyxoviridae The concept that proteolytic activation of a viral glycoprotein is an important factor in virus replication was first established with the fusion (F) protein of Sendai virus (Homma and Ohuchi 1973; Scheid and Choppin 1974). Cleavage of this protein results in the exposition of a highly conserved hydrophobic sequence at the amino terminus of the F1 fragment (Fig. 1) (Gething et al. 1978; Scheid et al. 1978) which induces fusion of the viral envelope with the plasma membrane of the target cell, thereby allowing entry of the viral genome into the cytoplasm. Cleavage of F is therefore necessary for virus infectivity. Evidence from various biochemical and biophysical data indicates that the membrane fusion characteristic of paramyxoviruses is driven by a hydrophobic interaction of the F1 amino terminus with the lipid bilayer...
Springer eBooks, 2018
The use of general descriptive names, registered names, trademarks, service marks, etc. in this p... more The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, express or implied, with respect to the material contained herein or for any errors or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
European journal of biochemistry, Sep 1, 1974
Virology, 1995
Inhibition of endoproteolytic cleavage of glycoprotein B (gB; gpUL55) of human cytomegalovirus wa... more Inhibition of endoproteolytic cleavage of glycoprotein B (gB; gpUL55) of human cytomegalovirus was achieved by treatment of infected fibroblasts with decanoyl peptidyl chloromethyl ketone (decRVKR-CMK), which inhibits the action of cellular subtilisin-like endoproteases with the amino acid recognition motif R X K/R R. Uncleaved gB precusor molecules of 160 kDa that were accumulated were endogiycosidase H resistant, suggesting that correct cellular transport occurred in the presence of the drug. The inhibitor also prevented endoproteolytic gB processing in CV-1 cells infected with a recombinant vaccinia virus-gB construct (WgB). Evidence for direct involvement of the ubiquitous subtilisin-like endoprotease furin in gB cleavage was obtained from the observation that coinfection of CV-1 cells with VVgB and a recombinant vaccinia-human furin construct reestablished endoproteolytic activity which was normally absent late after infection with WgB alone.
European journal of biochemistry, Nov 1, 1975
ABSTRACT
European journal of biochemistry, Dec 1, 1975
Biochemical Society Transactions, Apr 1, 1981
Gestational day-9.5 r a t conceptuses were cultured f o r 48h i n v i t r o i n glucose-and vitam... more Gestational day-9.5 r a t conceptuses were cultured f o r 48h i n v i t r o i n glucose-and vitamin-supplemented dialysed r a t serum. Embryos developed normally. When the serum p r o t e i n s were [3Hl leucine-labelled, t r i c h l o r o a c e t i c acid-insoluble r a d i o a c t i v i t y was incorporated i n t o embryo and yolk sac. Serum cont a i n i n g 1251-labelled polyvinylpyrrolidone o r
Biological Chemistry, 1998
... 1433 -1440, December 1998 · Copyright © by Walter de Gruyter · Berlin · New York Identificati... more ... 1433 -1440, December 1998 · Copyright © by Walter de Gruyter · Berlin · New York Identification and Characterization of Spodoptera frugiperda Furin: A Thermostable Subtilisin-Like Endoprotease Michael Cieplik3, Hans-Dieter Klenk and Wolfgang Garten* Institut für Virologie ...
Journal of Biological Chemistry, 1995
bioRxiv (Cold Spring Harbor Laboratory), Apr 15, 2020
Journal of Virology, 2010
Proteolytic cleavage of the influenza virus surface glycoprotein hemagglutinin (HA) by host cell ... more Proteolytic cleavage of the influenza virus surface glycoprotein hemagglutinin (HA) by host cell proteases is crucial for infectivity and virus spread. The proteases HAT (human airway trypsin-like protease) and TMPRSS2 (transmembrane protease serine S1 member 2) known to be present in the human airways were previously identified as proteases that cleave HA. We studied subcellular localization of HA cleavage and cleavage inhibition of seasonal influenza virus A/Memphis/14/96 (H1N1) and pandemic virus A/Hamburg/5/2009 (H1N1) in MDCK cells that express HAT and TMPRSS2 under doxycycline-induced transcriptional activation. We made the following observations: (i) HA is cleaved by membrane-bound TMPRSS2 and HAT and not by soluble forms released into the supernatant; (ii) HAT cleaves newly synthesized HA before or during the release of progeny virions and HA of incoming viruses prior to endocytosis at the cell surface, whereas TMPRSS2 cleaves newly synthesized HA within the cell and is not ...
Journal of Medicinal Chemistry, 2010
Furin belongs to the family of proprotein convertases (PCs) and is involved in numerous normal ph... more Furin belongs to the family of proprotein convertases (PCs) and is involved in numerous normal physiological and pathogenic processes, such as viral propagation, bacterial toxin activation, cancer, and metastasis. Furin and related furin-like PCs cleave their substrates at characteristic multibasic consensus sequences, preferentially after an arginine residue. By incorporating decarboxylated arginine mimetics in the P1 position of substrate analogue peptidic inhibitors, we could identify highly potent furin inhibitors. The most potent compound, phenylacetyl-Arg-Val-Arg-4-amidinobenzylamide (15), inhibits furin with a K i value of 0.81 nM and has also comparable affinity to other PCs like PC1/3, PACE4, and PC5/6, whereas PC2 and PC7 or trypsin-like serine proteases were poorly affected. In fowl plague virus (influenza A, H7N1)-infected MDCK cells, inhibitor 15 inhibited proteolytic hemagglutinin cleavage and was able to reduce virus propagation in a long-term infection test. Molecular modeling revealed several key interactions of the 4-amidinobenzylamide residue in the S1 pocket of furin contributing to the excellent affinity of these inhibitors.
Pathogenicity, i.e., the ability of a virus to induce disease in the infected organism, is the re... more Pathogenicity, i.e., the ability of a virus to induce disease in the infected organism, is the result of a complex interplay of a multitude of factors that are determined by the biological, biochemical, and genetic characteristics of the virus on the one hand, and the reactivity of the host on the other. Thus, a molecular basis for viral pathogenicity is not easy to define. It is reasonable, however, to assume that clinical disease becomes manifest, if cells of vital function are altered or killed by the infecting virus. Since the tropism of a virus for a host cell represents primarily an interaction between the surface components of the virus and receptors of the host cell, it is appealing to postulate that surface structures of a virus might determine pathogenicity. This concept is supported by recent results obtained with myxoviruses. It will be shown that the surface glycoproteins of these viruses acquire biological activity through proteolytic cleavage and that this activation ...
PubMed, Feb 9, 2000
Antibodies specific to the N-terminal 34-aminoacid peptide of the major nucleocapsid protein NP o... more Antibodies specific to the N-terminal 34-aminoacid peptide of the major nucleocapsid protein NP of human influenza A viruses were obtained. The NP proteolytic cleavage occurring in infected cells late in infection has been demonstrated using this antibody. The antibody specifically reacted with noncleaved NP of 56 kD m.w. but not with cleaved NP of 53 kD m.w. This indicates that NP56-->NP53 intracellular cleavage released the N-terminal 3 kD peptide of Np molecule. The released 3 kD peptide did not accumulate in infected cells. Since the RNA-binding and nucleus migrating signals are located in the N-terminus of NP molecule, presumably, the terminal cleavage is important for intracellular NP functions.
SUMMARY Processing of the hemagglutinin involves transport from the rough endoplasmic reticulum t... more SUMMARY Processing of the hemagglutinin involves transport from the rough endoplasmic reticulum to the plasma membrane, glycosylation, and proteolytic cleavage. The tight coupling of these events is demonstrated in experiments in which mutants of fowl plague virus (FPV) with a temperature sensitive defect in transport have been analyzed. Two groups of such mutants have been characterized. With the first group (ts1, ts227), the hemagglutinin is arrested in the rough endoplasmic reticulum. With the second group (ts482, 532, 651), the hemagglutinin migrates to the Golgi apparatus, but does not reach the plasma membrane. The distribution of the carbohydrate side chains on the FPV hemagglutinin has been elucidated. HA 1 has 4 type I side chains attached to asparagine residues 12, 28, 123, and 149. The potential attachment site at asparagine 231 is not glycosylated. HA2 has a type II chain at asparagine 406 and a type I chain at asparagine 478. Comparison with other hemagglutinins demonstrates that the side chains in positions 12, 28, and 478 are conserved. The cleavage site has been analyzed with H3, H7, and H10 hemagglutinins. The available evidence indicates that proteolytic activation involves the action of a trypsin-like protease followed by the action of an exopeptidase of the carboxypeptidase B type: After in vitro cleavage with trypsin, v^iich is paralleled by activation of infectivity, the N-terminus of HA 2 and presumably the C-terminus of HA. are identical to those obtained after in vivo cleavage. This indicates that the carboxypeptidase B activity is present in the virion. After cleavage with the non- activating enzymes thermolysin and chymo trypsin, the cleavage site is shifted by a few amino acids. These observations indicate that activation of infectivity requires a highly specific amino acid sequence at the cleavage site.
Cold Spring Harbor Monograph Archive, 1994
Endoproteolytic cleavage, usually at arginine residues, is a common posttranslational modificatio... more Endoproteolytic cleavage, usually at arginine residues, is a common posttranslational modification of membrane and secretory proteins on the exocytotic transport route. Such proteins include precursors of peptide hormones, neuropeptides, growth factors, coagulation factors, serum albumin, cell-surface receptors, and adhesion molecules. All of these proteins play important roles in a large variety of different biological processes, and these functions depend on proteolytic cleavage of the proteins (for review, see Barr 1991). The same type of processing has also been observed with many viral membrane proteins (Fig. 1) and, in some instances, proved to be a crucial factor in determining organ and host tropism, spread of infection, and pathogenicity of these viruses. VIRUSES WITH SPIKE PROTEINS UNDERGOING POSTTRANSLATIONAL PROTEOLYTIC CLEAVAGE Paramyxoviridae The concept that proteolytic activation of a viral glycoprotein is an important factor in virus replication was first established with the fusion (F) protein of Sendai virus (Homma and Ohuchi 1973; Scheid and Choppin 1974). Cleavage of this protein results in the exposition of a highly conserved hydrophobic sequence at the amino terminus of the F1 fragment (Fig. 1) (Gething et al. 1978; Scheid et al. 1978) which induces fusion of the viral envelope with the plasma membrane of the target cell, thereby allowing entry of the viral genome into the cytoplasm. Cleavage of F is therefore necessary for virus infectivity. Evidence from various biochemical and biophysical data indicates that the membrane fusion characteristic of paramyxoviruses is driven by a hydrophobic interaction of the F1 amino terminus with the lipid bilayer...
Springer eBooks, 2018
The use of general descriptive names, registered names, trademarks, service marks, etc. in this p... more The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, express or implied, with respect to the material contained herein or for any errors or omissions that may have been made. The publisher remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
European journal of biochemistry, Sep 1, 1974
Virology, 1995
Inhibition of endoproteolytic cleavage of glycoprotein B (gB; gpUL55) of human cytomegalovirus wa... more Inhibition of endoproteolytic cleavage of glycoprotein B (gB; gpUL55) of human cytomegalovirus was achieved by treatment of infected fibroblasts with decanoyl peptidyl chloromethyl ketone (decRVKR-CMK), which inhibits the action of cellular subtilisin-like endoproteases with the amino acid recognition motif R X K/R R. Uncleaved gB precusor molecules of 160 kDa that were accumulated were endogiycosidase H resistant, suggesting that correct cellular transport occurred in the presence of the drug. The inhibitor also prevented endoproteolytic gB processing in CV-1 cells infected with a recombinant vaccinia virus-gB construct (WgB). Evidence for direct involvement of the ubiquitous subtilisin-like endoprotease furin in gB cleavage was obtained from the observation that coinfection of CV-1 cells with VVgB and a recombinant vaccinia-human furin construct reestablished endoproteolytic activity which was normally absent late after infection with WgB alone.
European journal of biochemistry, Nov 1, 1975
ABSTRACT
European journal of biochemistry, Dec 1, 1975
Biochemical Society Transactions, Apr 1, 1981
Gestational day-9.5 r a t conceptuses were cultured f o r 48h i n v i t r o i n glucose-and vitam... more Gestational day-9.5 r a t conceptuses were cultured f o r 48h i n v i t r o i n glucose-and vitamin-supplemented dialysed r a t serum. Embryos developed normally. When the serum p r o t e i n s were [3Hl leucine-labelled, t r i c h l o r o a c e t i c acid-insoluble r a d i o a c t i v i t y was incorporated i n t o embryo and yolk sac. Serum cont a i n i n g 1251-labelled polyvinylpyrrolidone o r
Biological Chemistry, 1998
... 1433 -1440, December 1998 · Copyright © by Walter de Gruyter · Berlin · New York Identificati... more ... 1433 -1440, December 1998 · Copyright © by Walter de Gruyter · Berlin · New York Identification and Characterization of Spodoptera frugiperda Furin: A Thermostable Subtilisin-Like Endoprotease Michael Cieplik3, Hans-Dieter Klenk and Wolfgang Garten* Institut für Virologie ...
Journal of Biological Chemistry, 1995
bioRxiv (Cold Spring Harbor Laboratory), Apr 15, 2020
Journal of Virology, 2010
Proteolytic cleavage of the influenza virus surface glycoprotein hemagglutinin (HA) by host cell ... more Proteolytic cleavage of the influenza virus surface glycoprotein hemagglutinin (HA) by host cell proteases is crucial for infectivity and virus spread. The proteases HAT (human airway trypsin-like protease) and TMPRSS2 (transmembrane protease serine S1 member 2) known to be present in the human airways were previously identified as proteases that cleave HA. We studied subcellular localization of HA cleavage and cleavage inhibition of seasonal influenza virus A/Memphis/14/96 (H1N1) and pandemic virus A/Hamburg/5/2009 (H1N1) in MDCK cells that express HAT and TMPRSS2 under doxycycline-induced transcriptional activation. We made the following observations: (i) HA is cleaved by membrane-bound TMPRSS2 and HAT and not by soluble forms released into the supernatant; (ii) HAT cleaves newly synthesized HA before or during the release of progeny virions and HA of incoming viruses prior to endocytosis at the cell surface, whereas TMPRSS2 cleaves newly synthesized HA within the cell and is not ...
Journal of Medicinal Chemistry, 2010
Furin belongs to the family of proprotein convertases (PCs) and is involved in numerous normal ph... more Furin belongs to the family of proprotein convertases (PCs) and is involved in numerous normal physiological and pathogenic processes, such as viral propagation, bacterial toxin activation, cancer, and metastasis. Furin and related furin-like PCs cleave their substrates at characteristic multibasic consensus sequences, preferentially after an arginine residue. By incorporating decarboxylated arginine mimetics in the P1 position of substrate analogue peptidic inhibitors, we could identify highly potent furin inhibitors. The most potent compound, phenylacetyl-Arg-Val-Arg-4-amidinobenzylamide (15), inhibits furin with a K i value of 0.81 nM and has also comparable affinity to other PCs like PC1/3, PACE4, and PC5/6, whereas PC2 and PC7 or trypsin-like serine proteases were poorly affected. In fowl plague virus (influenza A, H7N1)-infected MDCK cells, inhibitor 15 inhibited proteolytic hemagglutinin cleavage and was able to reduce virus propagation in a long-term infection test. Molecular modeling revealed several key interactions of the 4-amidinobenzylamide residue in the S1 pocket of furin contributing to the excellent affinity of these inhibitors.
Pathogenicity, i.e., the ability of a virus to induce disease in the infected organism, is the re... more Pathogenicity, i.e., the ability of a virus to induce disease in the infected organism, is the result of a complex interplay of a multitude of factors that are determined by the biological, biochemical, and genetic characteristics of the virus on the one hand, and the reactivity of the host on the other. Thus, a molecular basis for viral pathogenicity is not easy to define. It is reasonable, however, to assume that clinical disease becomes manifest, if cells of vital function are altered or killed by the infecting virus. Since the tropism of a virus for a host cell represents primarily an interaction between the surface components of the virus and receptors of the host cell, it is appealing to postulate that surface structures of a virus might determine pathogenicity. This concept is supported by recent results obtained with myxoviruses. It will be shown that the surface glycoproteins of these viruses acquire biological activity through proteolytic cleavage and that this activation ...