Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.10.1 (ERK)
95,504 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have investigated a phosphatase-sensitive sequential epitope of the nucleoprotein (N), one of the phosphoproteins of rabies virus, which is recognized by the monoclonal antibody (MAb) #5-2-26. The epitope was shared in common by all of the rabies virus strains we tested, including the HEP, ERA, CVS and Japanese strains (Nishigahara and Komatsukawa). Thin layer chromatography of the acid hydrolyzates of 32P-labeled N protein showed that the protein contained phosphoserine and phosphothreonine at a molar ratio of about 4 to 1, while no phosphotyrosine was detected. Immunoprecipitation studies with several deletion mutants of the N protein showed that the epitope is located in a region spanning from amino acid 344 to 415. If the phosphatase-sensitive epitope is located at or near the phosphoamino acid, the location of the latter could be narrowed further to a region from amino acid 354 to 389 by comparing the amino-acid sequences among the viral strains. To examine this assumption, point mutation was introduced by amino-acid substitution with alanine at either of five potential phosphorylation sites (i.e., positions 354, 375, 377, 386 and 389) in the 354-389 region. Among those, only one substitution, at position 389, greatly affected the antigenicity. Substitution of serine-389 by threonine also reduced the antigenicity. These results strongly suggest that serine-389 is a phosphorylation site and essential for constructing or stabilizing the antigenic structure for MAb 5-2-26.
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PMID:Identification of a phosphatase-sensitive epitope of rabies virus nucleoprotein which is recognized by a monoclonal antibody 5-2-26. 913 Feb 35

To investigate the RNA polymerase of rabies virus, we cloned a cDNA of the catalytic subunit (called L protein because of its large molecular size) of the HEP-Flury strain, an avirulent strain obtained by high frequencies of serial embryonated hen egg passages. Nucleotide sequencing showed that the cDNA encodes a long polypeptide of 2,127 amino acids (Mr. 242,938). A comparison of the deduced amino acid sequence with that of other strains (PV and SAD B19) indicated that the sequence was highly conserved, except for several amino acid substitutions which were accumulated in some limited regions. A fragment of the cDNA was used for expression in Escherichia coli (E. coli) to prepare the L antigen for raising the antibodies in rabbits. Immunoprecipitation studies with the rabbit antiserum showed that the polypeptides produced in the L cDNA-transfected COS-7 cells displayed almost the same electrophoretic mobility as that of authentic L protein. Immunofluorescence studies indicated that both L and P (another subunit of RNA polymerase) proteins displayed colocalized distribution with the nucleocapsid antigen (N) in the cytoplasmic inclusion bodies, where envelope proteins (G and M) were absent. On the other hand, expression of the L protein alone did not cause inclusion body-like granular distribution, suggesting that the inclusion body-like accumulation depends on certain interaction(s) with other viral gene products, probably with the ribonucleoproteins comprising the inclusion bodies.
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PMID:Studies on rabies virus RNA polymerase: 1. cDNA cloning of the catalytic subunit (L protein) of avirulent HEP-flury strain and its expression in animal cells. 971 1

Acute infectious disease presentations among many at-risk patient groups (eg, uninsured, homeless, and recent immigrants) are frequently seen in emergency departments. Therefore EDs may be useful sentinel sites for infectious disease surveillance. This article describes the background, development, and implementation of EMERGE ncy ID NET, an interdisciplinary, multicenter, ED-based network for research of emerging infectious diseases. EMERGE ncy ID NET was established in cooperation with the National Center for Infectious Diseases, Centers for Disease Control and Prevention (CDC) as part of the CDC's strategy to expand and complement existing disease detection and control activities. The network is based at 11 university-affiliated, urban hospital EDs with a combined annual patient visit census of more than 900,000. Data are collected during ED evaluation of patients with specific clinical syndromes, and are electronically stored, transferred, and analyzed at a central receiving site. Current projects include investigation of bloody diarrhea and the prevalence of Shiga toxin-producing Escherichia coli, animal exposures and rabies postexposure prophylaxis practices, seizures and prevalence of neurocysticercosis, nosocomial ED Mycobacterium tuberculosis transmission, and hospital isolation bed use for adults admitted for pneumonia or suspected tuberculosis. EMERGE ncy ID NET also was developed to be a mechanism for rapidly responding to new diseases or epidemics. Future plans include study of antimicrobial use, meningitis, and encephalitis, and consideration of other public health concerns such as injury and national and international network expansion.
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PMID:EMERGEncy ID NET: an emergency department-based emerging infections sentinel network. The EMERGEncy ID NET Study Group. 983 68

To investigate the nature and intracellular behavior of the matrix (M) protein of an avirulent strain (HEP-Flury) of rabies virus, we cloned and sequenced the cDNA of the protein. Using expression vectors pZIP-NeoSV(X)1 and pCDM8, the cDNA was transfected to animal cells (BHK-21 and COS-7) with or without coexpression of viral glycoprotein (G). When M protein alone was expressed in the cells, it displayed homogeneous distribution in the whole cell including the nucleus. In contrast, coexpression with G protein resulted in the abolishment of nuclear distribution of M antigen, and both of the antigens displayed a colocalized distribution in the cell, especially at the cellular membrane as seen in the virus-infected cells, while the distribution of G antigen was not affected by coexpressed M antigen. Immunoprecipitation studies revealed that M protein was coprecipitated with G protein by anti-G antibody, and vice versa, although cross-linking with dithiobis(succinimidyl propionate) was necessary for coprecipitation because of their easier dissociation in the presence of sodium deoxycholate. These results suggest that M protein intimately associates with G protein, which may affect or regulate the behavior (e.g., intracellular localization) of M protein. Studies with deletion mutants of M protein indicate that an internal region around the amino acids from 115 to 151 is essential for the M protein to preserve its binding ability to G protein.
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PMID:Intracellular behavior of rabies virus matrix protein (M) is determined by the viral glycoprotein (G). 1033 96

We investigated a virus-neutralizing conformational epitope of the rabies virus glycoprotein (G) that is recognized by an anti-G monoclonal antibody (mAb; #1-46-12) and shared by most of the laboratory strains of the virus. To investigate the epitope structure, we isolated escape mutants from the HEP-Flury virus (wild-type; wt) after repeated passages in culture in the presence of the mAb. Immunofluorescence studies indicated that the mutants could be classified into two groups; the Group I lacked the epitope, while Group II preserved the epitope. The latter was dominant under the passage conditions, since Group I disappeared during the continuous passages. G proteins showed different electrophoretic mobilities; G protein of Group I migrated at the same rate as wt G protein, while that of Group II migrated at a slower rate, which was shown to be due to acquisition of an additional oligosaccharide side chain. Nucleotide sequencing of the G gene strongly suggested that amino acid substitutions at Thr-36 by Pro and Ser-39 by Thr of the G protein are responsible for the escape mutations of Groups I and II, respectively. The latter is a unique mutation of the rabies virus that allows the G protein to be glycosylated additionally at Asn-37, a potential glycosylation site that is not glycosylated in the parent virus, in preserving the epitope-positive conformation. These results suggest that to keep the 1-46-12 epitope structure is of greater survival advantage for the virus to escape the neutralization than to destroy it, which could be achieved by acquiring an additional oligosaccharide chain at Asn-37.
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PMID:Studies on the escape mutants of rabies virus which are resistant to neutralization by a highly conserved conformational epitope-specific monoclonal antibody #1-46-12. 1222 31

Monoclonal antibody (mAb) #1-30-44 recognized an acid-sensitive conformational epitope of rabies virus glycoprotein (G). The antigenicity of G protein exposed on the cell surface was lost when the infected cells were exposed to pH 5.8. By comparing the deduced amino acid sequence of G protein between the HEP-Flury strain and the epitope-negative CVS strain as well as the mAb-resistant escape mutants, two distant sites that contained Lys-202 and Asn-336 were shown to be involved in the epitope formation. Lys-202 is located in the so-called neurotoxin-like sequence, while Asn-336 is included in antigenic site III and is very near the amino acid at position 333, which is known to affect greatly the neuropathogenicity of rabies virus when changed. Consistent with this finding, antigenicity of a neurovirulent revertant of the HEP-Flury strain, in which Gln-333 of G protein was replaced by Arg, was also affected as shown by its greatly decreased reactivity with mAb #1-30-44 compared to that of the original avirulent HEP virus. Based on these results, we hypothesize that the neurotoxin-like domain and some amino acids in antigenic site III come into contact with each other to form a conformational epitope for mAb #1-30-44, and such a configuration would be lost when exposed to acidic conditions to perform a certain low pH-dependent function of G protein.
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PMID:Mapping of the low pH-sensitive conformational epitope of rabies virus glycoprotein recognized by a monoclonal antibody #1-30-44. 1295 44

The authors report on an investigation into recent advances in rabies treatment designed to assess the various claims put forward, their relative merits, and whether they hold true when Indian strains of street virus are used for challenge. The results are presented of treatment with antirabies serum alone, with 5% Semple vaccine, with a combination of serum and vaccination, and with HEP Flury vaccine with and without serum.Antirabies serum alone, given before or after infection, while definitely prolonging the incubation period, had no saving effect. Serum seemed to confer some protection when it was obtained from animals of the same species which had survived an infection with the particular challenge virus.With doses of 5% Semple vaccine comparable to those administered to human beings it was possible to confer solid protection to animals against virulent strains of street virus provided the treatment was started 7 days before challenge.Combined therapy with serum and vaccine given after infection was of great value under certain conditions. There appeared to be an optimum relationship between the quantity of serum given and the antigenicity and dosage of vaccine administered.HEP Flury vaccine given before infection gave very good protection. But serum and HEP Flury vaccine administered after infection was not of value against an Indian strain of street virus.
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PMID:Advances in rabies treatment; an experimental evaluation. 1351 Nov 41

The authors have compared the value of multiple doses of duck-embryo and HEP Flury vaccine with that of pooled 5% sheep-brain vaccine in experimental rabies infection in guinea-pigs. They found that the duck-embryo vaccine given in a dosage corresponding to 14 ml of 10% vaccine (the dosage recommended for human treatment), either alone or with antirabies serum, gave no protection and that, even when administered in a dosage corresponding to 140 ml of 5% pooled vaccine, both the duck-embryo and the HEP Flury vaccines, whether alone or with serum, conferred little protection. Pooled phenolized vaccine under identical conditions gave good results. The immunogenicity of duck-embryo and HEP Flury vaccines, given before infection, was also inferior to that of pooled vaccine; and the duck-embryo vaccine was found to be a poorer antigen than the pooled vaccine in mouse potency tests.The authors conclude that the dosage of duck-embryo vaccine recommended for human treatment is inadequate and that the HEP Flury vaccine in its present form is unsuitable for post-infection treatment.
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PMID:THE VALUE OF DUCK-EMBRYO VACCINE AND HIGH-EGG-PASSAGE FLURY VACCINE IN EXPERIMENTAL RABIES INFECTION IN GUINEA-PIGS. 1406 70

To obviate the foreign protein reactions experienced with the use of hyperimmune serum in rabies-exposed individuals, an attempt was made to produce a rabies antiserum of human origin.Five doses of an inactivated rabies virus duck-egg vaccine were administered to 34 volunteers at 4-day intervals (i.e., on days 0, 4, 8, 12 and 16). An additional dose of chick-embryo attenuated virus vaccine-Flury HEP (high egg passage)-was given on the 46th day, followed by a final booster dose of duck-egg vaccine on the 288th day. Twenty-four days later, i.e., on the 312th day after the first dose, the participants were bled and the serum pooled and converted to gamma globulin.These volunteers, having no initial antibody, responded with variable titres, the pooled serum having a titre of 1: 100 against 50 LD(50) of rabies virus in neutralization tests and the gamma globulin prepared from this pool a titre of 1: 300.In five individuals inoculated with the antirabies gamma globulin, blood samples tested at intervals for residual antibody showed significant titres through 21 days.While the passive antibody levels resulting from the administration of a more potent immune horse serum were much higher than those achieved by the weaker human antirabies gamma globulin used, the decrease in titre was more gradual with the human globulin. With more booster inoculations in a larger group of human volunteers, it is believed that a human rabies immune gamma globulin could be produced which would be equal in effect to immune horse serum. The advantages of a human source of antibody in rabies prophylaxis are discussed.
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PMID:Human antirabies gamma globulin. 1440 20

Rabies virus (RV) deficient in the P gene was generated by reverse genetics from cDNA of HEP-Flury strain lacking the entire P gene. The defective virus was propagated and amplified by rescue of virus, using a cell line that complemented the functions of the deficient gene. The P gene-deficient (def-P) virus replicated its genome and produced progeny viruses in the cell lines that constitutively expressed the P protein, although it grew at a slightly retarded rate compared to the parental strain. In contrast, no progeny virus was produced in the infected host when the def-P virus-infected cells that did not express the P protein. However, we found that the def-P virus had the ability to perform primary transcription (by the virion-associated polymerase) in the infected host without de novo P protein synthesis. The def-P virus was apathogenic in adult and suckling mice, even when inoculated intracranially. Inoculation of def-P virus in mice induced high levels of virus-neutralizing antibody (VNA) and conferred protective immunity against a lethal rabies infection. These results demonstrate the potential utility of gene-deficient virus as a novel live attenuated rabies vaccine.
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PMID:Generation and characterization of P gene-deficient rabies virus. 1497 55


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