scholarly journals Influenza C Virus CM2 Protein Is Produced from a 374-Amino-Acid Protein (P42) by Signal Peptidase Cleavage

1999 ◽  
Vol 73 (1) ◽  
pp. 46-50 ◽  
Author(s):  
Seiji Hongo ◽  
Kanetsu Sugawara ◽  
Yasushi Muraki ◽  
Yoko Matsuzaki ◽  
Emi Takashita ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Matrix Protein ◽  
Signal Peptidase ◽  
M Gene ◽  
Recognition Motif ◽  
Acid Protein ◽  
Influenza C Virus ◽  
Amino Acid Protein

ABSTRACT Although unspliced mRNA from influenza C virus RNA segment 6 (M gene) has a single open reading frame capable of encoding a 374-amino-acid protein (Mr, 42,000), the major polypeptide synthesized from this mRNA species is the CM2 protein, with an Mr of 18,000. The present study was performed to investigate the mechanism by which CM2 is generated from the unspliced mRNA. It was reported previously that the 374-amino-acid protein (P42) is an integral membrane protein having two internal hydrophobic domains, one of which (residues 241 to 252) is followed by two sequences (252 Ile-Thr-Ser and 257 Ala-Ser-Ala) favorable for cleavage by signal peptidase. To examine the possibility that P42 is cleaved by signal peptidase after Ser residue 254 or Ala residue 259 to yield CM2, we constructed three mutated M gene cDNAs in which either or both of the two sequences were eliminated and tested their ability to synthesize CM2 in the transfected COS cells. The results showed that CM2 synthesis was blocked completely when the second recognition motif for signal peptidase was removed. It was also found that when the mRNA transcript of the wild-type M gene was translated in vitro, P42, but not CM2, was synthesized in the absence of dog pancreas microsomal membranes, whereas CM2, in addition to a polypeptide (designated M1′) slightly larger than matrix protein (M1), was synthesized in the presence of microsomes. When the same experiment was done with the transcript of the mutated M gene in which the second recognition motif was removed, synthesis of CM2 could not be seen, even in the presence of microsomes. From these results, we conclude that cleavage of P42 by signal peptidase after Ala residue 259 produces CM2, composed of the C-terminal 115 amino acids, in addition to M1′, composed of the N-terminal 259 amino acids.

scholarly journals Genetic Organization and Mode of Action of a Novel Bacteriocin, Bacteriocin 51: Determinant of VanA-Type Vancomycin-Resistant Enterococcus faecium

2011 ◽  
Vol 55 (9) ◽  
pp. 4352-4360 ◽  
Author(s):  
Hitoshi Yamashita ◽  
Haruyoshi Tomita ◽  
Takako Inoue ◽  
Yasuyoshi Ike
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Enterococcus Faecium ◽  
Promoter Sequence ◽  
Signal Peptidase ◽  
Start Codon ◽  
Content Type ◽  
Acid Protein ◽  
Vancomycin Resistant ◽  
Amino Acid Protein

ABSTRACTBacteriocin 51 (Bac 51) is encoded on the mobile plasmid pHY (6,037 bp), which was isolated from vancomycin-resistantEnterococcus faeciumVRE38. Bacteriocin 51 is active againstE. faecium,E. hirae, andE. durans. Sequence analysis of pHY showed that it encodes nine open reading frames (ORFs) from ORF1 to ORF9 (in that order). Genetic analysis suggested that ORF1 and ORF2, which were designatedbacAandbacB, respectively, are the bacteriocin and immunity genes.bacAencodes a 144-amino-acid protein. The deduced BacA protein has a typical signal sequence at its amino terminus, and a potential signal peptidase-processing site corresponding to the V-E-A sequence is located between the 37th and 39th amino acids. The predicted mature BacA protein consists of 105 amino acids. A potential promoter sequence was identified upstream of the start codon.bacBencodes a 55-amino-acid protein. No obvious promoter or terminator sequence was identified betweenbacAandbacB. Northern blot analysis ofbacAandbacBwith abacARNA probe produced a transcript of approximately 700 nucleotides, which corresponded to the combined nucleotide sizes ofbacAandbacB, indicating that transcription was initiated from the promoter upstream ofbacA, continued throughbacB, and was terminated at the terminator downstream ofbacB. The transcription start site was determined to be the T nucleotide located 6 nucleotides downstream from the −10 promoter sequence. These results indicate thatbacAandbacBconstitute an operon and thatbacAis the bacteriocin structural gene whilebacBis the immunity gene. The purified C-terminally His tagged BacA protein of Bac 51 showed bacteriostatic activity against the indicator strain. The purified C-terminally His tagged BacA protein of Bac 32 (whose mature BacA protein has 54 amino acids) and the culture filtrates of the Bac 31- and Bac 43-producingE. faecalisstrain FA2-2 showed bactericidal activity. Bac 31 and Bac 43 are pore-forming bacteriocins, unlike the newly characterized bacteriocin Bac 51.

scholarly journals LASS3 (longevity assurance homologue 3) is a mainly testis-specific (dihydro)ceramide synthase with relatively broad substrate specificity

2006 ◽  
Vol 398 (3) ◽  
pp. 531-538 ◽  
Author(s):  
Yukiko Mizutani ◽  
Akio Kihara ◽  
Yasuyuki Igarashi
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Family Members ◽  
Fatty Acyl ◽  
Ceramide Synthase ◽  
Broad Substrate Specificity ◽  
Acid Protein ◽  
Amino Acid Protein

The LASS (longevity assurance homologue) family members are highly conserved from yeasts to mammals. Five mouse and human LASS family members, namely LASS1, LASS2, LASS4, LASS5 and LASS6, have been identified and characterized. In the present study we cloned two transcriptional variants of hitherto-uncharacterized mouse LASS3 cDNA, which encode a 384-amino-acid protein (LASS3) and a 419-amino-acid protein (LASS3-long). In vivo, [3H]dihydrosphingosine labelling and electrospray-ionization MS revealed that overproduction of either LASS3 isoform results in increases in several ceramide species, with some preference toward those having middle- to long-chain-fatty acyl-CoAs. A similar substrate preference was observed in an in vitro (dihydro)ceramide synthase assay. These results indicate that LASS3 possesses (dihydro)ceramide synthesis activity with relatively broad substrate specificity. We also found that, except for a weak display in skin, LASS3 mRNA expression is limited almost solely to testis, implying that LASS3 plays an important role in this gland.

scholarly journals Identification of a 374 amino acid protein encoded by RNA segment 6 of influenza C virus.

1998 ◽  
Vol 79 (9) ◽  
pp. 2207-2213 ◽  
Author(s):  
S Hongo ◽  
K Sugawara ◽  
Y Matsuzaki ◽  
K Nakamura ◽  
F Kitame ◽  
...  
Keyword(s):  
Amino Acid ◽  
Acid Protein ◽  
Influenza C Virus ◽  
Amino Acid Protein

Analysis of amino acids, proteins, carbohydrates and lipids in food by capillary electromigration methods: a review

2016 ◽  
Vol 8 (18) ◽  
pp. 3649-3680 ◽  
Author(s):  
Marcone A. L. de Oliveira ◽  
Brenda L. S. Porto ◽  
Carina de A. Bastos ◽  
Céphora M. Sabarense ◽  
Fernando A. S. Vaz ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Food Samples ◽  
Carbohydrate Analysis ◽  
Acid Protein ◽  
Amino Acid Protein

We review the literature covering the evolution of amino acid, protein, lipid and carbohydrate analysis in food samples by electromigration techniques over the last 20 years.

Characterization of the small endogenous plasmid of Halobacterium strain SB3 and its use in transformation of H. halobium

1989 ◽  
Vol 35 (1) ◽  
pp. 86-91 ◽  
Author(s):  
Neil R. Hackett ◽  
Shiladitya DasSarma
Keyword(s):  
Amino Acid ◽  
Nucleotide Sequence ◽  
Molecular Biology ◽  
Base Pair ◽  
Copy Number ◽  
Halobacterium Halobium ◽  
Acid Protein ◽  
Amino Acid Protein

To study the molecular biology of the halophilic archaebacterium Halobacterium halobium, the introduction of DNA engineered in vitro is desirable. As a first step in developing a cloning vector, the complete 1736 base pair nucleotide sequence of the natural, high copy number, Halobacterium plasmid pHSB1 has been determined. The plasmid was found to show homology to the small plasmids of Halobacterium strains GRB and GN101. Plasmid pHSB1 encodes a 317 amino acid protein of unknown function. The related halophile, H. halobium, could be transformed by pHSB1, demonstrating its utility as the basis of a cloning vector.Key words: archaebacteria, Halobacterium, plasmid.

scholarly journals Assembly of flagellar radial spoke proteins in Chlamydomonas: identification of the axoneme binding domain of radial spoke protein 3.

1993 ◽  
Vol 123 (1) ◽  
pp. 183-190 ◽  
Author(s):  
D R Diener ◽  
L H Ang ◽  
J L Rosenbaum
Keyword(s):  
Amino Acids ◽  
Binding Activity ◽  
Binding Domain ◽  
Flagellar Motility ◽  
Radial Spoke ◽  
Acid Protein ◽  
Radial Spokes ◽  
Carboxy Terminal ◽  
Amino Acid Protein

Radial spokes of the eukaryotic flagellum extend from the A tubule of each outer doublet microtubule toward the central pair microtubules. In the paralyzed flagella mutant of Chlamydomonas pf14, a mutation in the gene for one of 17 polypeptides that comprise the radial spokes results in flagella that lack all 17 spoke components. The defective gene product, radial spoke protein 3 (RSP3), is, therefore, pivotal to the assembly of the entire spoke and may attach the spoke to the axoneme. We have synthesized RSP3 in vitro and assayed its binding to axonemes from pf14 cells to determine if RSP3 can attach to spokeless axonemes. In vitro, RSP3 binds to pf14 axonemes, but not to wild-type axonemes or microtubules polymerized from purified chick brain tubulin. The sole axoneme binding domain of RSP3 is located within amino acids 1-85 of the 516 amino acid protein; deletion of these amino acids abolishes binding by RSP3. Fusion of amino acids 1-85 or 42-85 to an unrelated protein confers complete or partial binding activity, respectively, to the fusion protein. Transformation of pf14 cells with mutagenized RSP3 genes indicates that amino acids 18-87 of RSP3 are important to its function, but that the carboxy-terminal 140 amino acids can be deleted with little effect on radial spoke assembly or flagellar motility.

scholarly journals A Metabonomics Investigation into the Therapeutic Effects of BuChang NaoXinTong Capsules on Reversing the Amino Acid-Protein Interaction Network of Cerebral Ischemia

2019 ◽  
Vol 2019 ◽  
pp. 1-14 ◽  
Author(s):  
Jing Xu ◽  
Xin Liu ◽  
Liyu Luo ◽  
Liying Tang ◽  
Na Guo ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Cerebral Ischemia ◽  
Protein Interaction ◽  
System Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Acid Decarboxylase ◽  
Therapeutic Effects ◽  
Acid Protein ◽  
Amino Acid Protein

Background. Amino acids (AAs) in cerebrospinal fluid (CSF) play a pivotal role in cerebral ischemia (CI). BuChang NaoXinTong Capsules (BNC) are widely prescribed in Chinese medicine for the treatment of cerebrovascular and cardiovascular diseases. Methods. In order to investigate the therapeutic effects and pharmacological mechanisms of BNC on reversing CI from a system level, an amino acid-protein interaction imbalanced network of CI containing metabolites of AAs, key regulatory enzymes, and proteins was constructed for the first time. Furthermore, a novel method for detecting the ten AAs in CSF was developed by UPLC-QQQ-MS in an effort to validate the imbalanced networks and the therapeutic effects of BNC via analysis of metabolites. Results. Based on a middle cerebral artery occlusion (MCAO) rat model, the dynamic levels of amino acids in CSF 3, 6, 12, and 24 h after MCAO were analyzed. Up to 24 h, the accumulated nine AA biomarkers were found to significantly change in the MCAO group compared to the sham group and exhibited an obvious tendency for returning to baseline values after BNC treatment. In addition, based on the imbalanced network of CI, four key enzymes that regulate the generation of BNC-mediated AA biomarkers were selected and validated using an enzyme-linked immunosorbent assay and western blotting. Finally, aromatic-L-amino-acid decarboxylase (AADC) was found to be one of the putative targets for BNC-mediated protection against CI. Conclusion. This study provides new strategies to explore the mechanism of cerebral ischemia and help discover the potential mechanism of BNC.

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