The deoxyribonucleoprotein complex from the archaebacterium Methanosarcina barkeri: characterization and ultrastructural localization of the chromosomal protein MC1

1988 ◽  
Vol 34 (8) ◽  
pp. 931-937 ◽  
Author(s):  
Marlène Imbert ◽  
Bernard Laine ◽  
Gérard Prensier ◽  
Jean-Pierre Touzel ◽  
Pierre Sautière
Keyword(s):  
Protein A ◽  
Ultrastructural Localization ◽  
Methanosarcina Barkeri ◽  
Chromosomal Protein ◽  
Amino Acid Residues ◽  
Total Proteins ◽  
Nuclease Digestion ◽  
Basic Polypeptide ◽  
Deoxyribonucleoprotein Complex

The Methanosarcina barkeri protein, methanogen chromosomal protein 1 (MC1), a basic polypeptide of 93 amino acid residues, has been localized in the DNA-rich areas on immunolabelled bacterial cryosections revealed with the protein A – gold method. This localization strongly supports the previous studies concerning the association of the protein MC1 with the DNA in vivo. In the M. barkeri deoxyribonucleoprotein complex, the protein MC1 accounts for about 90% of the total proteins, and the protein MC1 so DNA ratio (by weight) is equal to 0.11. Staphylococcal nuclease digestion data and microscopic observations have failed to reveal the presence of repeating structural units and suggest that the protein MC1 is unevenly distributed along the DNA.

The Primary Structure of Horseshoe Crab (Tachypleus tridentatus) Coagulogen and its Homologies with Platelet Factor 4

1979 ◽  
Author(s):  
T. Takagi ◽  
S. Nakamura ◽  
Y. Hokama ◽  
T. Miyata ◽  
M. Niwa ◽  
...  
Keyword(s):  
Amino Acid ◽  
Horseshoe Crab ◽  
Protein A ◽  
Platelet Factor 4 ◽  
Amino Acid Residues ◽  
Platelet Factor ◽  
Disulfide Linkages ◽  
A Chain ◽  
Basic Polypeptide ◽  
Large Peptide

Horseshoe crab coagulogen consists of a single basic polypeptide chain having the total 175 amino acid residues. Upon gelation of this protein, a large peptide, named peptide C, which has 28 amino acid residues, was released, and the resulting gel protein consisted of A and B chains, bridged by two disulfide linkages. The sequence studies on peptide C, A chain and B chain provided that their sequences have a significant homology partly with primate fibrinopeptide B and largely with human platelet factor 4. The facts suggest that these are derived from a common ancester or that the coagulogen is a prototype of platelet factor 4. The whole sequence of coagulogen was as follows:Ala-Asp-Thr-Asn-Ala-Pro-Ile-Cys-Leu-Cys-Asp-Glu-Pro-Gly-Val-Leu-Gly-Arg-Thr-Gln-Ile-Val-Thr-Thr-Glu-Ile-Lys-Asp-Lys-Ue-Glu-Lys-Ala-Val-Clu-Ala-Val-Ala-Gln-Glu-Ser-Gly-Val-Ser-Gly-Arg-Cly-Fhe-Ser-Ile-Phe-Ser-His-His-Pro-Val-Phe-Arg-Glu-Cys-Gly-Lys-Tyr-Glu-Cys-Arg-Thr-Val-Arg-Fro-Glu-Hls-Ser-Arg-Cys-Tyr-Asn-Phe-Pro-Pro-Phe-Thr-His-Phe-Lys-Leu-Glu-Cys-Pro-Val-Ser-Thr-Arg-Asp-Cys-Glu-Pro-Va1-Phe-Gly-Tyr-Thr-Va1-Ala-Gly-Glu-Phe-Arg-Va1-1le-Va1-GIn-Ala-Pro-Arg-Ala-Cly-Phe-Arg-GIn-Cys-Val-Trp-Cin-His-Lys-Cys-Arg-Phe-Giy-Ser-Asn-Ser-Cys-Gly-Tyr-Asn-Gly-Arg-Cys-Thr-Gln-Gln-Arg-Ser-Val-Val-Arg-Leu-Val-Thr-Tyr-Asn-Leu-Clu-Lys-Asp-Cly-Phe-Leu-Cys-Glu-Ser-Phe-Arg-Thr-Cys-Gys-Cly-Cys-Pro-Cys-Arg-Ser-Phe,

The Amino Acid Sequence Studies On Limulus Polyphemus Coagulogen

1981 ◽  
Author(s):  
T Miyata ◽  
M Umezu ◽  
S Iwanaga
Keyword(s):  
Amino Acid ◽  
Horseshoe Crab ◽  
Polypeptide Chain ◽  
Protein A ◽  
Limulus Polyphemus ◽  
Amino Acid Residues ◽  
Sequence Determination ◽  
Disulfide Linkages ◽  
Basic Polypeptide ◽  
Large Peptide

Coagulogen, which is a clottable protein found in the amebocyte lysate of Limulus polyphemus, consists of a single basic polypeptide chain having the total of 174 amino acid residues. Upon gelation of this protein, a large peptide, named peptide C, which has 28 amino acid residues, was released, and the resulting gel protein consisted of A and B chains, bridged by two disulfide linkages. This work is to determine the whole sequence of B chain and to compare with the primary structure of Tachypleus tridentatus (Japanese horseshoe crab) coagulogen previously established. The Limulus B chain was digested with trypsin and the resulting fragments were separated into 17 major peptides. For the sequence determination, manual Edman degradation was employed and resulting PTH amino acids were identified by HPLC. These analyses established the sequence of the isolated tryptic peptides, ending with the COOH-terminal serine, and the following whole sequence of Limulus coagulogen was presumed, comparing with that of Tachypleus coagulogen.Tachypleus: NH2-A-D-T-N-A-P-I-C-L-C-D-E-P-G-V-L-G-R-T-Q-I-V- Limulus : NH2-G-D-P-N-V-P-T-C-L-C-E-E-P-T-L-L-G-R-K-V-I-V- T-T-E-I-K-D-K-I-E-K-A-V-E-A-V-A-Q-E-S-G-V-S-G-R-G-F-S-I-F-S- S-Q-E-T-K-D-K-I-E-E-A-V-Q-A-I-T-B-K-D-E-I-S-G-R⋅G-F-S-I-F-G⋅ H-H-P-V-F-R-E-C-G-K-Y-E-C-R-T-V-R-P-E-H-S-R-C-Y-N-F-P-P-F-T- H-G-P-A-F-K-E-C-G-K⋅Y-E-C-R⋅T-V-T-S-E-D-S-R⋅C-Y-N-F-F-P-F-S⋅ H-F-K-L-E-C-P-V-S-T-R-D-C-E-P-V-F-G-Y-T-V-A-G-E-F-R-V-I-V-Q- H⋅F⋅H⋅P⋅Z⋅C⋅P⋅T⋅S⋅T⋅S⋅B⋅C⋅Z⋅P⋅V⋅L⋅S⋅Y⋅T⋅V⋅A⋅G⋅Z⋅F⋅R⋅I-I-V-Q- A-P-R-A-G-F-R-Q-C-V-W-Q-H-K-C-R-F-G-S-N-S-C-G-Y-N-G-R-C-T-Q- A-P-K.A-G-F-R⋅D-L-E(W)V⋅H⋅K⋅C-R⋅A-Y-G-S-N-F-C-Q-S-Z-R⋅C-T-Q- Q-R-S-V-V-R-L-V-T-Y-N-L-E-K-D-G-F-L-C-E-S-F-R-T-C-C-G-C-P-C- Q-R-S-V-V-R-L-V-T-Y-D-L-E-K⋅ G-V-F-F-C-E-N-V-R-T-C-C-G-C-P-C- R-S-F-COOH R-S-C00H

scholarly journals Novel 30 kDa protein possessing ATP-binding and chaperone activities

1997 ◽  
Vol 326 (2) ◽  
pp. 567-572 ◽  
Author(s):  
Hideaki ITOH ◽  
Yohtalou TASHIMA
Keyword(s):  
Amino Acid ◽  
Divalent Cations ◽  
Protein A ◽  
Amino Acid Sequences ◽  
Stable Complex ◽  
Dependent Manner ◽  
Amino Acid Residues ◽  
Protection Effect ◽  
Α Helix

A 30 kDa protein was purified from pig liver cytosol by using ATP-Sepharose and Green A column chromatography. The partial amino acid sequences of the protein (95 amino acid residues) had no similarity with any proteins recorded in data banks. The protein was able to form a stable complex with unfolded dihydrofolate reductase (DHFR). The spontaneous refolding of chemically denatured DHFR was arrested by the 30 kDa protein. This inhibition presumably results from the formation of a stable complex between the 30 kDa protein and DHFR. Bound DHFR could be released from the protein with ATP. The protein also showed protease resistance in an ATP-dependent manner. Incubation of the 30 kDa protein with 5 mM ATP resulted in its resistance to V8 protease or to trypsin treated with 1-chloro-4-phenyl-3-L-toluene-p-sulphonamidobutan-2-one. Divalent cations enhanced the ATP-protection effect. CD analysis of the 30 kDa protein showed that ATP induced an increase in the β-pleated sheet content and a decrease in the α-helix content of the 30 kDa protein. These results suggest that the 30 kDa protein, a novel cytosolic protein, might have an affinity for ATP, a chaperonin activity, and and an ATP-protection effect against some proteases in vivo.

Oxytocin analogues with non-coded amino acid residues in position 8: [8-Neopentylglycine]oxytocin and [8-cycloleucine]oxytocin

1987 ◽  
Vol 52 (9) ◽  
pp. 2317-2325 ◽  
Author(s):  
Jan Hlaváček ◽  
Jan Pospíšek ◽  
Jiřina Slaninová ◽  
Walter Y. Chan ◽  
Victor J. Hruby
Keyword(s):  
Amino Acid ◽  
Solid Phase Synthesis ◽  
Solid Phase ◽  
The Other ◽  
Amino Acid Residues ◽  
Phase Synthesis ◽  
Other Hand ◽  
Fragment Condensation

[8-Neopentylglycine]oxytocin (II) and [8-cycloleucine]oxytocin (III) were prepared by a combination of solid-phase synthesis and fragment condensation. Both analogues exhibited decreased uterotonic potency in vitro, each being about 15-30% that of oxytocin. Analogue II also displayed similarly decreased uterotonic potency in vivo and galactogogic potency. On the other hand, analogue III exhibited almost the same potency as oxytocin in the uterotonic assay in vivo and in the galactogogic assay.

Synthesis of Fragments of the Vasoactive Intestinal Peptide (VIP) and Analysis of Their Residual Biological Activities

1995 ◽  
Vol 60 (7) ◽  
pp. 1229-1235 ◽  
Author(s):  
Ivana Zoulíková ◽  
Ivan Svoboda ◽  
Jiří Velek ◽  
Václav Kašička ◽  
Jiřina Slaninová ◽  
...  
Keyword(s):  
Amino Acid ◽  
Biological Activities ◽  
Residual Activity ◽  
Detailed Examination ◽  
Amino Acid Residues ◽  
Linear Peptide ◽  
Zone Electrophoresis ◽  
Capillary Zone

The vasoactive intestinal (poly)peptide (VIP) is a linear peptide containing 28 amino acid residues, whose primary structure indicates a low metabolic stability. The following VIP fragments, as potential metabolites, and their analogues were prepared by synthesis on a solid: [His(Dnp)1]VIP(1-10), VIP(11-14), [D-Arg12]VIP(11-14), [Lys(Pac)15,21,Arg20]VIP(15-22), and VIP(23-28). After purification, the peptides were characterized by amino acid analysis, mass spectrometry, RP HPLC, and capillary zone electrophoresis. In some tests, detailed examination of the biological activity of the substances in vivo and in vitro gave evidence of a low, residual activity of some fragments, viz. a depressoric activity in vivo for [His(Dnp)1]VIP(1-10) and a stimulating activity for the release of α-amylase in vitro and in vivo for [Lys(Pac)15,21,Arg20]VIP(15-22) and VIP(23-28).

scholarly journals Characterization of the SARS-CoV-2 coronavirus X4-like accessory protein

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Olanrewaju Ayodeji Durojaye ◽  
Nkwachukwu Oziamara Okoro ◽  
Arome Solomon Odiba
Keyword(s):  
Rapid Development ◽  
Protein A ◽  
The Novel ◽  
Quality Model ◽  
A Value ◽  
Model Protein ◽  
Novel Coronavirus ◽  
Global Threat

Abstract Background The novel coronavirus SARS-CoV-2 is currently a global threat to health and economies. Therapeutics and vaccines are in rapid development; however, none of these therapeutics are considered as absolute cure, and the potential to mutate makes it necessary to find therapeutics that target a highly conserved regions of the viral structure. Results In this study, we characterized an essential but poorly understood coronavirus accessory X4 protein, a core and stable component of the SARS-CoV family. Sequence analysis shows a conserved ~ 90% identity between the SARS-CoV-2 and previously characterized X4 protein in the database. QMEAN Z score of the model protein shows a value of around 0.5, within the acceptable range 0–1. A MolProbity score of 2.96 was obtained for the model protein and indicates a good quality model. The model has Ramachandran values of φ = − 57o and ψ = − 47o for α-helices and values of φ = − 130o and ψ = + 140o for twisted sheets. Conclusions The protein data obtained from this study provides robust information for further in vitro and in vivo experiment, targeted at devising therapeutics against the virus. Phylogenetic analysis further supports previous evidence that the SARS-CoV-2 is positioned with the SL-CoVZC45, BtRs-BetaCoV/YN2018B and the RS4231 Bat SARS-like corona viruses.

scholarly journals The Yeast Recombinational Repair Protein Rad59 Interacts With Rad52 and Stimulates Single-Strand Annealing

Genetics ◽  
2001 ◽  
Vol 159 (2) ◽  
pp. 515-525 ◽  
Author(s):  
Allison P Davis ◽  
Lorraine S Symington
Keyword(s):  
Protein A ◽  
Single Strand ◽  
Physical Interaction ◽  
Dna Double Strand Breaks ◽  
Single Stranded Dna ◽  
Single Strand Annealing ◽  
Strand Annealing ◽  
Recombination Pathway

Abstract The yeast RAD52 gene is essential for homology-dependent repair of DNA double-strand breaks. In vitro, Rad52 binds to single- and double-stranded DNA and promotes annealing of complementary single-stranded DNA. Genetic studies indicate that the Rad52 and Rad59 proteins act in the same recombination pathway either as a complex or through overlapping functions. Here we demonstrate physical interaction between Rad52 and Rad59 using the yeast two-hybrid system and co-immunoprecipitation from yeast extracts. Purified Rad59 efficiently anneals complementary oligonucleotides and is able to overcome the inhibition to annealing imposed by replication protein A (RPA). Although Rad59 has strand-annealing activity by itself in vitro, this activity is insufficient to promote strand annealing in vivo in the absence of Rad52. The rfa1-D288Y allele partially suppresses the in vivo strand-annealing defect of rad52 mutants, but this is independent of RAD59. These results suggest that in vivo Rad59 is unable to compete with RPA for single-stranded DNA and therefore is unable to promote single-strand annealing. Instead, Rad59 appears to augment the activity of Rad52 in strand annealing.

scholarly journals Brown Spiders’ Phospholipases-D with Potential Therapeutic Applications: Functional Assessment of Mutant Isoforms

Biomedicines ◽  
2021 ◽  
Vol 9 (3) ◽  
pp. 320
Author(s):  
Thaís Pereira da Silva ◽  
Fernando Jacomini de Castro ◽  
Larissa Vuitika ◽  
Nayanne Louise Costacurta Polli ◽  
Bruno César Antunes ◽  
...  
Keyword(s):  
Structural Changes ◽  
Capillary Permeability ◽  
Structural Data ◽  
Histopathological Analysis ◽  
Amino Acid Residues ◽  
Wild Type ◽  
Antigenic Properties ◽  
Harmful Effects

Phospholipases-D (PLDs) found in Loxosceles spiders’ venoms are responsible for the dermonecrosis triggered by envenomation. PLDs can also induce other local and systemic effects, such as massive inflammatory response, edema, and hemolysis. Recombinant PLDs reproduce all of the deleterious effects induced by Loxosceles whole venoms. Herein, wild type and mutant PLDs of two species involved in accidents—L. gaucho and L. laeta—were recombinantly expressed and characterized. The mutations are related to amino acid residues relevant for catalysis (H12-H47), magnesium ion coordination (E32-D34) and binding to phospholipid substrates (Y228 and Y228-Y229-W230). Circular dichroism and structural data demonstrated that the mutant isoforms did not undergo significant structural changes. Immunoassays showed that mutant PLDs exhibit conserved epitopes and kept their antigenic properties despite the mutations. Both in vitro (sphingomyelinase activity and hemolysis) and in vivo (capillary permeability, dermonecrotic activity, and histopathological analysis) assays showed that the PLDs with mutations H12-H47, E32-D34, and Y228-Y229-W230 displayed only residual activities. Results indicate that these mutant toxins are suitable for use as antigens to obtain neutralizing antisera with enhanced properties since they will be based on the most deleterious toxins in the venom and without causing severe harmful effects to the animals in which these sera are produced.

scholarly journals UV-induced Hyperphosphorylation of Replication Protein A Depends on DNA Replication and Expression of ATM Protein

2001 ◽  
Vol 12 (5) ◽  
pp. 1199-1213 ◽  
Author(s):  
Gregory G. Oakley ◽  
Lisa I. Loberg ◽  
Jiaqin Yao ◽  
Mary A. Risinger ◽  
Remy L. Yunker ◽  
...  
Keyword(s):  
Dna Replication ◽  
Uv Radiation ◽  
Excision Repair ◽  
Genetic Disorder ◽  
Protein A ◽  
Dna Recombination ◽  
Delayed Response ◽  
Replication Intermediates

Exposure to DNA-damaging agents triggers signal transduction pathways that are thought to play a role in maintenance of genomic stability. A key protein in the cellular processes of nucleotide excision repair, DNA recombination, and DNA double-strand break repair is the single-stranded DNA binding protein, RPA. We showed previously that the p34 subunit of RPA becomes hyperphosphorylated as a delayed response (4–8 h) to UV radiation (10–30 J/m2). Here we show that UV-induced RPA-p34 hyperphosphorylation depends on expression of ATM, the product of the gene mutated in the human genetic disorder ataxia telangiectasia (A-T). UV-induced RPA-p34 hyperphosphorylation was not observed in A-T cells, but this response was restored by ATM expression. Furthermore, purified ATM kinase phosphorylates the p34 subunit of RPA complex in vitro at many of the same sites that are phosphorylated in vivo after UV radiation. Induction of this DNA damage response was also dependent on DNA replication; inhibition of DNA replication by aphidicolin prevented induction of RPA-p34 hyperphosphorylation by UV radiation. We postulate that this pathway is triggered by the accumulation of aberrant DNA replication intermediates, resulting from DNA replication fork blockage by UV photoproducts. Further, we suggest that RPA-p34 is hyperphosphorylated as a participant in the recombinational postreplication repair of these replication products. Successful resolution of these replication intermediates reduces the accumulation of chromosomal aberrations that would otherwise occur as a consequence of UV radiation.

Diffusible amyloid oligomers trigger systemic amyloidosis in mice

2008 ◽  
Vol 415 (2) ◽  
pp. 207-215 ◽  
Author(s):  
Sivanesan Senthilkumar ◽  
Edwin Chang ◽  
Rajadas Jayakumar
Keyword(s):  
Protein A ◽  
Mouse Tissue ◽  
Cytokine Induction ◽  
Amyloidogenic Proteins ◽  
Enhancing Factor ◽  
Amyloid Oligomers ◽  
Apparent Relationship ◽  
Amyloid Enhancing Factor ◽  
Local Accumulation

AA (amyloid protein A) amyloidosis in mice is markedly accelerated when the animals are given, in addition to an inflammatory stimulus, an intravenous injection of protein extracted from AA-laden mouse tissue. Previous findings affirm that AA fibrils can enhance the in vivo amyloidogenic process by a nucleation seeding mechanism. Accumulating evidence suggests that globular aggregates rather than fibrils are the toxic entities responsible for cell death. In the present study we report on structural and morphological features of AEF (amyloid-enhancing factor), a compound extracted and partially purified from amyloid-laden spleen. Surprisingly, the chief amyloidogenic material identified in the active AEF was diffusible globular oligomers. This partially purified active extract triggered amyloid deposition in vital organs when injected intravenously into mice. This implies that such a phenomenon could have been inflicted through the nucleation seeding potential of toxic oligomers in association with altered cytokine induction. In the present study we report an apparent relationship between altered cytokine expression and AA accumulation in systemically inflamed tissues. The prevalence of serum AA monomers and proteolytic oligomers in spleen AEF is consistent to suggest that extrahepatic serum AA processing might lead to local accumulation of amyloidogenic proteins at the serum AA production site.

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