The Protein Encoded at the 3′ End of the Serine Protease Gene of Aeromonas sobria Functions as a Chaperone in the Production of the Protease

ABSTRACT For the successful production of Aeromonas sobria serine protease (ASP), open reading frame 2 (ORF2) protein, encoded at the 3′ end of the protease operon, is required. In this study, we examined the action of ORF2 protein. The results showed that the protein associated with ASP in the periplasm and helped ASP to form an active structure.

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Anti-Tumor Effects of a Newly Cloned Serine Protease from the Marine Annelid, Arenicola cristata

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.998-999.183 ◽

2014 ◽

Vol 998-999 ◽

pp. 183-186

Author(s):

Chun Ling Zhao ◽

Wen Jing Yu ◽

Ji Yu Ju

Keyword(s):

Amino Acids ◽

Serine Protease ◽

Protease Gene ◽

Mature Peptide ◽

Reading Frame ◽

Conserved Sequence ◽

Arenicola Cristata ◽

Serine Protease Family ◽

Rapid Amplification ◽

Serine Protease Gene

We cloned a new serine protease gene from the marine annelid,Arenicola cristataby rapid amplification of cDNA ends. The full-length cDNA of 901bp contained an open reading frame of 774bp encoding 258 amino acids. Sequence analysis of the deduced amino acids indicated that this protease belonged to serine protease family and contained highly conserved sequence GDSGGP. An expression vector, harboring the mature peptide ofArenicola cristataprotease, was constructed and transformed intoE.coli. The purified recombinant protein could inhibit proliferation of cancer cells in a dose-dependant way and induce apoptosis. These results indicated that the recombinant protease ofArenicola cristata, as a new member of serine protease family, might be valuable in developing anti-tumor agents.

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Antigenic Domains of the Open Reading Frame 2-Encoded Protein of Hepatitis E Virus

Journal of Clinical Microbiology ◽

10.1128/jcm.37.9.2863-2871.1999 ◽

1999 ◽

Vol 37 (9) ◽

pp. 2863-2871 ◽

Cited By ~ 38

Author(s):

Yury E. Khudyakov ◽

Elena N. Lopareva ◽

Danny L. Jue ◽

Tamara K. Crews ◽

S. P. Thyagarajan ◽

...

Keyword(s):

Synthetic Peptides ◽

Hepatitis E Virus ◽

Hepatitis E ◽

Open Reading Frame ◽

Reading Frame ◽

Orf2 Protein ◽

Positive Serum ◽

Antigenic Domains ◽

Open Reading Frame 2 ◽

Antigenic Epitopes

The antigenic composition of the hepatitis E virus (HEV) protein encoded by open reading frame 2 (ORF2) was determined by using synthetic peptides. Three sets of overlapping 18-, 25-, and 30-mer peptides, with each set spanning the entire ORF2 protein of the HEV Burma strain, were synthesized. All synthetic peptides were tested by enzyme immunoassay against a panel of 32 anti-HEV-positive serum specimens obtained from acutely HEV-infected persons. Six antigenic domains within the ORF2 protein were identified. Domains 1 and 6 located at the N and C termini of the ORF2 protein, respectively, contain strong immunoglobulin G (IgG) and IgM antigenic epitopes that can be efficiently modeled with peptides of different sizes. In contrast, antigenic epitopes identified within the two central domains (3 and 4) were modeled more efficiently with 30-mer peptides than with either 18- or 25-mers. Domain 2 located at amino acids (aa) 143 to 222 was modeled best with 25-mer peptides. A few 30-mer synthetic peptides derived from domain 5 identified at aa 490 to 579 demonstrated strong IgM antigenic reactivity. Several 30-mer synthetic peptides derived from domains 1, 4, and 6 immunoreacted with IgG or IgM with more than 70% of anti-HEV-positive serum specimens. Thus, the results of this study demonstrate the existence of six diagnostically relevant antigenic domains within the HEV ORF2 protein.

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Antigenic Characterization of ORF2 and ORF3 Proteins of Hepatitis E Virus (HEV)

Viruses ◽

10.3390/v13071385 ◽

2021 ◽

Vol 13 (7) ◽

pp. 1385

Author(s):

Giulia Pezzoni ◽

Lidia Stercoli ◽

Eleonora Pegoiani ◽

Emiliana Brocchi

Keyword(s):

Hepatitis E Virus ◽

Hepatitis E ◽

Recombinant Antigen ◽

Open Reading Frame ◽

Reading Frame ◽

E Coli ◽

Antigenic Characterization ◽

Antigenic Properties ◽

Open Reading Frame 2

To evaluate the antigenic properties of Hepatitis E Virus (HEV) Open Reading Frame 2 and 3 (ORF2 and ORF3) codified proteins, we expressed different portions of ORF2 and the entire ORF3 in E. coli, a truncated ORF2, was also expressed in baculovirus. A panel of 37 monoclonal antibodies (MAbs) was raised against ORF2 (1–660 amino acids) and MAbs were mapped and characterized using the ORF2 expressed portions. Selected HEV positive and negative swine sera were used to evaluate ORF2 and ORF3 antigens’ immunogenicity. The MAbs were clustered in six groups identifying six antigenic regions along the ORF2. Only MAbs binding to the sixth ORF2 antigenic region (394–608 aa) were found to compete with HEV positive sera and efficiently catch the recombinant antigen expressed in baculovirus. The ORF2 portion from 394–608 aa demonstrated to include most immunogenic epitopes with 85% of HEV positive swine sera reacting against the region from 461–544 aa. Only 5% of the selected HEV sera reacted against the ORF3 antigen.

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Expression and activity-dependent changes of a novel limbic-serine protease gene in the hippocampus

Journal of Neuroscience ◽

10.1523/jneurosci.15-07-05088.1995 ◽

1995 ◽

Vol 15 (7) ◽

pp. 5088-5097 ◽

Cited By ~ 151

Author(s):

ZL Chen ◽

S Yoshida ◽

K Kato ◽

Y Momota ◽

J Suzuki ◽

...

Keyword(s):

Serine Protease ◽

Protease Gene ◽

Activity Dependent ◽

Serine Protease Gene ◽

Expression And Activity

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Regulation of Metalloprotease Gene Expression in Vibrio vulnificus by a Vibrio harveyi LuxR Homologue

Journal of Bacteriology ◽

10.1128/jb.183.4.1369-1375.2001 ◽

2001 ◽

Vol 183 (4) ◽

pp. 1369-1375 ◽

Cited By ~ 74

Author(s):

Chung-Ping Shao ◽

Lien-I Hor

Keyword(s):

Parent Strain ◽

Vibrio Vulnificus ◽

Vibrio Harveyi ◽

Negative Regulation ◽

Open Reading Frame ◽

Protease Gene ◽

Reading Frame ◽

Allelic Exchange ◽

Exchange Technique

ABSTRACT Expression of the Vibrio vulnificus metalloprotease gene, vvp, was turned up rapidly when bacterial growth reached the late log phase. A similar pattern of expression has been found in the metalloprotease gene of Vibrio cholerae, and this has been shown to be regulated by a Vibrio harveyiLuxR-like transcriptional activator. To find out whether a LuxR homologue exists in V. vulnificus, a gene library of this organism was screened by colony hybridization using a probe derived from a sequence that is conserved in various luxR-like genes of vibrios. A gene containing a 618-bp open reading frame was identified and found to be identical to the smcR gene ofV. vulnificus reported previously. An isogenic SmcR-deficient (RD) mutant was further constructed by an in vivo allelic exchange technique. This mutant exhibited an extremely low level of vvp transcription compared with that of the parent strain. On the other hand, the cytolysin gene, vvhA, was expressed at a higher level in the RD mutant than in the parent strain during the log phase of growth. These data suggested that SmcR might not only be a positive regulator of the protease gene but might also be involved in negative regulation of the cytolysin gene. Virulence of the RD mutant in either normal or iron-overloaded mice challenged by intraperitoneal injection was comparable to that of the parent strain, indicating that SmcR is not required for V. vulnificusvirulence in mice.

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