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Tytuł:
Acetylation of methyl 5-amino-1H-[1,2,4]triazole-3-carboxylate
Autorzy:
Dżygiel, Anetta
MAsiukiewicz, Elżbieta
Rzeszotarska, Barbara
Powiązania:
https://bibliotekanauki.pl/articles/1044073.pdf
Data publikacji:
2001
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
hetareneamino acid
5-amino-1H-[1,2,4]triazole-3-carboxylate
Opis:
Acetylation with acetic anhydride of methyl 5-amino-1H-[1,2,4]triazole-3-carboxylate, one of the hetareneamino acids, was studied using HPLC,1H NMR, FTIR and GC-MS. The compound has a significantly decreased susceptibility to acetylation compared to 5-amino-1H-[1,2,4]triazole itself. Two isomeric diacetylated products were found.
Źródło:
Acta Biochimica Polonica; 2001, 48, 4; 1169-1173
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Application of 1H and 31P NMR to topological description of a model of biological membrane fusion Topological description of a model of biological membrane fusion
Autorzy:
Janiak-Osajca, Agnieszka
Timoszyk, Anna
Powiązania:
https://bibliotekanauki.pl/articles/1039735.pdf
Data publikacji:
2012
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
vesicle
topology
fusion
31P-NMR
1H-NMR
Opis:
The process of biological membrane fusion can be analysed by topological methods. Mathematical analysis of the fusion process of vesicles indicated two significant facts: the formation of an inner, transient structure (hexagonal phase - HII) and a translocation of some lipids within the membrane. This shift had a vector character and only occurred from the outer to the inner layer. Model membrane composed of phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylserine (PS) was studied. 31P- and 1H-NMR methods were used to describe the process of fusion. 31P-NMR spectra of multilamellar vesicles (MLV) were taken at various temperatures and concentrations of Ca2+ ions (natural fusiogenic agent). A 31P-NMR spectrum with the characteristic shape of the HII phase was obtained for the molar Ca2+/PS ratio of 2.0. During the study, 1H-NMR and 31P-NMR spectra for small unilamellar vesicle (SUV), which were dependent on time (concentration of Pr3+ ions was constant), were also recorded. The presence of the paramagnetic Pr3+ ions permits observation of separate signals from the hydrophilic part of the inner and outer lipid bilayers. The obtained results suggest that in the process of fusion translocation of phospholipid molecules takes place from the outer to the inner layer of the vesicle and size of the vesicles increase. The NMR study has showed that the intermediate state of the fusion process caused by Ca2+ ions is the HII phase. The experimental results obtained are in agreement with the topological model as well.
Źródło:
Acta Biochimica Polonica; 2012, 59, 2; 219-224
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Changes in kinesin expression in the CNS of mice with dynein heavy chain 1 mutation
Autorzy:
Kuźma-Kozakiewicz, Magdalena
Kaźmierczak, Beata
Usarek, Ewa
Barańczyk-Kuźma, Anna
Powiązania:
https://bibliotekanauki.pl/articles/1039605.pdf
Data publikacji:
2013
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Dync1h1 mutation
kinesins
Cra1/+ mice
expression
central nervous system
Cra1/SOD1 mice
Opis:
Dysfunction of fast axonal transport, vital for motor neurons, may lead to neurodegeneration. Anterograde transport is mediated by N-kinesins (KIFs), while retrograde transport by dynein 1 and, to a minor extent, by C-kinesins. In our earlier studies we observed changes in expression of N- and C-kinesins (KIF5A, 5C, C2) in G93ASOD1-linked mouse model of motor neuron degeneration. In the present work we analyze the profile of expression of the same kinesins in mice with a dynein 1 heavy chain mutation (Dync1h1, called Cra1), presenting similar clinical symptoms, and in Cra1/SOD1 mice with milder disease progression than SOD1 transgenics. We found significantly higher levels of mRNA for KIF5A and KIF5C but not the KIFC2 in the frontal cortex of symptomatic Cra1/+ mice (aged 365 days) compared to the wild-type controls. No changes in kinesin expression were found in the spinal cord of any age group and only mild changes in the hippocampus. The expression of kinesins in the cerebellum of the presymptomatic and symptomatic mice (aged 140 and 365 days, respectively) was much lower than in age-matched controls. In Cra1/SOD1 mice the changes in KIFs expression were similar or more severe than in the Cra1/+ groups, and they also appeared in the spinal cord. Thus, in mice with the Dync1h1 mutation, which impairs dynein 1-dependent retrograde transport, expression of kinesin mRNA is affected in various structures of the CNS and the changes are similar or milder than in mice with double Dync1h1/hSOD1G93A mutations.
Źródło:
Acta Biochimica Polonica; 2013, 60, 1; 51-55
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Detection of acylglycines in urine by 1H and 13C NMR for the diagnosis of inborn metabolic diseases
Autorzy:
Gryff-Keller, Adam
Kraska-Dziadecka, Anna
Kubica, Dominika
Powiązania:
https://bibliotekanauki.pl/articles/1039667.pdf
Data publikacji:
2012
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
13C NMR
urinalysis
inherited metabolic disease
1H NMR
acylglycine
Opis:
A range of inborn metabolic diseases result in abnormal accumulation of acylglycines in body fluids. Therefore, detection of these metabolites is important for diagnostic purposes. 1H and 13C NMR spectroscopies have successfully been applied for both qualitative and quantitative determinations of various acylglycines in urine samples from patients suffering from metabolic diseases connected with excretion of these compounds. Various acylglycines were identified in test urine samples from 15 patients suffering from five different metabolic diseases, providing information which could be crucial for their diagnoses. The paper reports complete 1H and 13C NMR data of 11 acylglycines, which is essential for this type of NMR analysis of body fluids. NMR spectroscopy has been proven effective in determining the presence as well as the levels of acylglycines in urine. The proposed method is rapid, simple and requires minimal sample treatment.
Źródło:
Acta Biochimica Polonica; 2012, 59, 4; 613-617
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Investigation of a wide spectrum of inherited metabolic disorders by 13C NMR spectroscopy
Autorzy:
Bal, Dominika
Kraska-Dziadecka, Anna
Gradowska, Wanda
Gryff-Keller, Adam
Powiązania:
https://bibliotekanauki.pl/articles/1040823.pdf
Data publikacji:
2008
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
urine
13C NMR
inherited metabolic disease
1H NMR
marker metabolite
Opis:
High-resolution 1H NMR spectroscopy of body fluids has proved to be very useful in diagnostics of inherited metabolic diseases, whereas 13C NMR remains almost unexploited. In this paper the application of 13C NMR spectroscopy of fivefold concentrated urine samples for diagnosis of selected metabolic diseases is reported. Various marker metabolites were identified in test urine samples from 33 patients suffering from 10 different diseases, providing information which could be crucial for their diagnoses. Spectra were accumulated for 2 h or overnight when using spectrometers operating at 9.4 or 4.7 T magnetic fields, respectively. Interpretation of the measurement results was based on a comparison of the peak positions in the measured spectrum with reference data. The paper contains a table with 13C NMR chemical shifts of 73 standard compounds. The method can be applied individually or as an auxiliary technique to 1H NMR or any other analytical method.
Źródło:
Acta Biochimica Polonica; 2008, 55, 1; 107-118
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
1H-Benzimidazole derivatives as mammalian DNA topoisomerase I inhibitors
Autorzy:
Alpan, A
Gunes, H
Topcu, Zeki
Powiązania:
https://bibliotekanauki.pl/articles/1040939.pdf
Data publikacji:
2007
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
1H-benzimidazole derivatives
type I DNA topoisomerase
plasmid supercoil relaxation assays
Opis:
Benzimidazole is one of the most important heterocyclic groups manifesting various biological properties, such as antibacterial, antifungal, antimicrobial, antiprotozoal and antihelmintic activities. Several benzimidazole derivatives are also active as inhibitors of type I DNA topoisomerases. In this study, three 1H-benzimidazole derivatives with different electronic characteristics at position 5-, namely 5-chloro-4-(1H-benzimidazole-2-yl)phenol (Cpd I), 5-methyl-4-(1H-benzimidazole-2-yl)phenol (Cpd II) and 4-(1H-benzimidazole-2-yl)phenol (Cpd III), were synthesized and evaluated for their effects on mammalian type I DNA topoisomerase activity using quantitative in vitro plasmid supercoil relaxation assays. For the structure elucidation of the compounds, melting points, UV, IR, 1H NMR, 13C NMR, mass spectral data and elemental analyses were interpreted. Among the compounds, 5-methyl-4-(1H-benzimidazole-2-yl)phenol (Cpd II) manifested relatively potent topoisomerase I inhibition.
Źródło:
Acta Biochimica Polonica; 2007, 54, 3; 561-565
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Differences in glutathione S-transferase pi expression in transgenic mice with symptoms of neurodegeneration
Autorzy:
Kaźmierczak, Beata
Kuźma-Kozakiewicz, Magdalena
Usarek, Ewa
Barańczyk-Kuźma, Anna
Powiązania:
https://bibliotekanauki.pl/articles/1039866.pdf
Data publikacji:
2011
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
motor neuron disease
Dync1h1 mutation
SOD1G93A mutation
central nervous system
transgenic mice
glutathione S-transferase pi
Opis:
Glutathione S-transferase pi (GST pi) is an enzyme involved in cell protection against toxic electrophiles and products of oxidative stress. GST pi expression was studied in transgenic mice hybrids (B6-C3H) with symptoms of neurodegeneration harboring SOD1G93A (SOD1/+), Dync1h1 (Cra1/+) and double (Cra1/SOD1) mutations, at presymptomatic and symptomatic stages (age 70, 140, 365 days) using RT-PCR and Western blotting. The main changes in GST pi expression were observed in mice with the SODG93A mutation. In SOD1/+ and Cra1/SOD1 transgenics, with the exception of cerebellum, the changes in GST pi-mRNA accompanied those in GST pi protein. In brain cortex of both groups the expression was unchanged at the presymptomatic (age 70 days) but was lower at the symptomatic stage (age 140 days) and at both stages in hippocampus and spinal cord of SOD1/+ but not of Cra1/SOD1 mice compared to age-matched wild-type controls. In cerebellum of the presymptomatic and the symptomatic SOD1/+ mice and presymptomatic Cra1/SOD1 mice, the GST pi-mRNA was drastically elevated but the protein level remained unchanged. In Cra1/+ transgenics there were no changes in GST pi expression in any CNS region both on the mRNA and on the protein level. It can be concluded that the SOD1G93A but not the Dync1h1 mutation significantly decreases detoxification efficiency of GST pi in CNS, however the Dync1h1 mutation reduces the effects caused by the SOD1G93A mutation. Despite similarities in neurological symptoms, the differences in GST pi expression between SOD1/+ and Cra1/+ transgenics indicate a distinct pathogenic entity of these two conditions.
Źródło:
Acta Biochimica Polonica; 2011, 58, 4; 621-626
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Translocation of polysialic acid across model membranes: Kinetic analysis and dynamic.
Autorzy:
Janas, Teresa
Krajiński, Henryk
Timoszyk, Anna
Janas, Tadeusz
Powiązania:
https://bibliotekanauki.pl/articles/1044181.pdf
Data publikacji:
2001
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
polysialic acid
1H-NMR
electrical equivalent circuit
polycation flux
polyanion flux
lipid bilayers
membrane transport
Opis:
Transmembrane translocation of polyion homopolymers takes place in the case of polyanionic polysialic acid (polySia), polyanionic polynucleotides and polycationic polypeptides. The purpose of this work was to determine the role of membrane electrical parameters on the kinetics of polyion translocation, the influence of polysialic acid on ion adsorption on positively charged membrane surface and the dynamics of the phospholipid hydrocarbon chains and choline group by using 1H-NMR. The analysis of polyion translocation was performed by using the electrical equivalent circuit of the membrane for the initial membrane potential equal to zero. The changes in polysialic acid flux was up to 75% after 1 ms in comparison with the zero-time flux. Both a decrease of membrane conductance and an increase of polyion chain length resulted in the diminution of this effect. An increase of praseodymium ions adsorption to positively charged liposomes and an increase of the rate of segmental movement of the -CH2 and -CH3 groups, and the choline headgrup of lipid molecules, was observed in the presence of polySia. The results show that the direction of the vectorial polyion translocation depends both on the membrane electrical properties and the degree of polymerization of the polymer, and that polysialic acid can modulate the degree of ion adsorption and the dynamics of membrane lipids.
Źródło:
Acta Biochimica Polonica; 2001, 48, 1; 163-173
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Self-association of Chaetopterus variopedatus sperm histone H1-like. Relevance of arginine content and possible physiological role
Autorzy:
Salvati, Daniela
Conforti, Salvatore
Conte, Mariachiara
Matassa, Danilo
Fucci, Laura
Piscopo, Marina
Powiązania:
https://bibliotekanauki.pl/articles/1040674.pdf
Data publikacji:
2008
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Chaetopterus variopedatus
arginine
sperm histone H1-like
Opis:
Self-association of histones H1 from calf thymus and from sperm of the marine worm Chaetopterus variopedatus was studied on native and glutaraldehyde cross-linked molecules by PAGE and by salt-induced turbidity measurements. Multiple polymers were generated by native sperm histone H1-like after glutaraldehyde cross-linking while the same treatment on its lysine- or arginine-modified derivatives and on somatic histone H1 failed to induce polymerization. This result suggests the relevance of arginine content in the formation of histone H1-like polymers particularly because Chaetopterus variopedatus and calf thymus histones H1 have similar content of lysine but different K/R ratio (2 and 15, respectively). Salt-induced turbidity experiments confirmed the high tendency of sperm histone H1-like to form oligomers, particularly in the presence of phosphate ions. Native PAGE analysis in the presence of phosphate supported this hypothesis. The reported results suggest that phosphate ions connecting lysine and arginine side chain groups contribute to the interaction of sperm histone H1-like with DNA in chromatin and play a key role in organization and stabilization of the chromatin higher order structures.
Źródło:
Acta Biochimica Polonica; 2008, 55, 4; 701-706
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Avian influenza in Poland
Autorzy:
Śmietanka, Krzysztof
Minta, Zenon
Powiązania:
https://bibliotekanauki.pl/articles/1039245.pdf
Data publikacji:
2014
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
avian influenza
H5N1 virus
Opis:
Poland has experienced four episodes of avian influenza (AI) outbreaks over the past two decades. The first epidemic was caused by a low pathogenicity (LPAIV) H7N7 subtype and occurred in fattening and breeder turkeys in 1995. Two waves of H5N1 high pathogenicity avian influenza (HPAI) took place in 2006 and 2007. In spring 2006, 64 cases of the H5N1 virus were detected, mostly in mute swans. In December 2007, ten outbreaks of H5N1 HPAI were detected in commercial poultry (n=9) and wild birds kept in captivity (n=1). The outbreaks in 2006 and 2007 were caused by genetically similar but clearly distinguishable viruses of the 2.2 clade. In 2013, an H9N2 avian influenza virus was detected in 4 fattening turkey holdings. The virus was low pathogenic and a phylogenetic study has shown a close relatedness to the Eurasian lineage of AIV of the wild bird origin. Neither preventive nor prophylactic vaccinations have ever been used in poultry or other birds. Emergency vaccinations using autogenous vaccine were introduced only to control the H7N7 LPAI outbreaks in 1995. The baseline surveillance for AI in live migratory birds and poultry provides a valuable insight into the ecology of AIV at the wild and domestic bird interface. Passive surveillance is in place of early detection of HPAIV infection in dead or moribund birds.
Źródło:
Acta Biochimica Polonica; 2014, 61, 3; 453-457
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
A clinical utility of a strip test for influenza A/B and comparison with detection by RT PCR
Autorzy:
Miarka, Maciej
Horban, Andrzej
Maliszewska, Henryka
Biliński, Przemysław
Prus-Kowalczuk, Wanda
Powiązania:
https://bibliotekanauki.pl/articles/1039251.pdf
Data publikacji:
2014
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
A/H1N1/v influenza virus
strip test
influenza RT PCR
Opis:
Introduction and objective: In June 2009 the World Health Organization announced influenza pandemic caused by A/H1N1/v virus. It became crucial to recognize new cases of A/H1N1/v infection. An effective screening diagnostic procedure was needed for patients suffering from influenza-like symptoms for making an initial diagnosis and analyzing epidemiological pattern of infection. We used a strip test for influenza A/B as a screening diagnostic procedure for patients suffering from influenza-like symptoms for making an initial diagnosis. For comparison, RT PCR for detecting A/H1N1/v was performed. The aim of this study was to assess the efficacy and sensitivity of the strip test and its value for making initial diagnosis of influenza A/H1N1/v. Material and methods: Strip testing for the influenza A/B infection was performed on 1123 patients with influenza-like symptoms in the Admission Unit of the Regional Infectious Diseases Hospital in Warsaw. Strip test results were analyzed according to the age of patients and season of the year. For 97 patients strip test results for detecting A/H1N1 infection were compared with those obtained by RT PCR. Results: There were no statistically significant differences found between the methods and strip testing demonstrated sensitivity of 61% and specificity of 71%. Conclusions: No statistically significant differences were found between the two methods, however, strip test had low sensitivity and specificity.
Źródło:
Acta Biochimica Polonica; 2014, 61, 3; 485-487
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
On the possibility that H1 histone interaction with DNA occurs through phosphates connecting lysine and arginine side chain groups
Autorzy:
Piscopo, Marina
De Petrocellis, Luciano
Conte, Mariachiara
Pulcrano, Giovanna
Geraci, Giuseppe
Powiązania:
https://bibliotekanauki.pl/articles/1041206.pdf
Data publikacji:
2006
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Chaetopterus variopedatus
protein structure
sperm H1 histone
arginine
lysine
ionic interaction
Opis:
Gel filtration and velocity of sedimentation analyses on native and on lysine- and arginine- modified forms of the annelid worm Chaetopterus variopedatus sperm H1 histone indicate that anion-mediated lysine - arginine interactions play a relevant role in the stabilization of the oligomeric states of the molecule. CD spectroscopy shows that phosphate anions are at least an order of magnitude more efficient than chloride as negatively charged groups connecting H1 lysines and arginines. Acetylation of lysines, although not altering grossly the H1 properties, causes a tenfold decrease of the structuring efficiency of phosphates. This suggests that DNA phosphates may be sandwiched between lysine and arginine groups of H1 histone when this molecule binds to chromatin, constituting a relevant parameter for the reciprocal stabilization of the protein and of the chromatin higher order structures.
Źródło:
Acta Biochimica Polonica; 2006, 53, 3; 507-513
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Antibody response to DNA vaccine against H5N1 avian influenza virus in broilers immunized according to three schedules
Autorzy:
Stachyra, Anna
Góra-Sochacka, Anna
Zagórski-Ostoja, Włodzimierz
Król, Ewelina
Sirko, Agnieszka
Powiązania:
https://bibliotekanauki.pl/articles/1039267.pdf
Data publikacji:
2014
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Avian influenza
H5N1
DNA vaccine
broilers
Opis:
Broiler type chickens were immunized intramuscularly with a DNA vaccine encoding hemagglutinin (HA) from H5N1 avian influenza virus. The chickens were divided into four groups: control group which was not immunized, a group which obtained only one dose, and two groups which were immunized twice, one group with a boost two weeks after the priming and the other four weeks. Blood samples were collected at several time points and the dynamics of the humoral response to the vaccine was studied. High level of anti-HA antibodies was detected only in the last two groups, that is in chickens immunized according to the prime-boost strategy, regardless of the schedule. An additional interesting observation of this study was detection of the cross-reactivity of an anti-H5 HA positive serum with H5N2 and H1N1 viruses, suggesting that the DNA vaccine tested can induce antibodies of a broad specificity.
Źródło:
Acta Biochimica Polonica; 2014, 61, 3; 593-596
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Possible computational filter to detect proteins associated to influenza A subtype H1N1
Autorzy:
Polanco, Carlos
Buhse, Thomas
Castañón-González, Jorge
Samaniego, José
Powiązania:
https://bibliotekanauki.pl/articles/1039197.pdf
Data publikacji:
2014
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Polarity Index Method
influenza A virus subtype H1N1
drug design
QSAR method
Opis:
The design of drugs with bioinformatics methods to identify proteins and peptides with a specific toxic action is increasingly recurrent. Here, we identify toxic proteins towards the influenza A virus subtype H1N1 located at the UniProt database. Our quantitative structure-activity relationship (QSAR) approach is based on the analysis of the linear peptide sequence with the so-called Polarity Index Method that shows an efficiency of 90% for proteins from the Uniprot Database. This method was exhaustively verified with the APD2, CPPsite, Uniprot, and AmyPDB databases as well as with the set of antibacterial peptides studied by del Rio et al. and Oldfield et al.
Źródło:
Acta Biochimica Polonica; 2014, 61, 4; 693-698
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
Tytuł:
Expression, purification and characterization of glycosylated influenza H5N1 hemagglutinin produced in Pichia pastoris
Autorzy:
Kopera, Edyta
Dwornyk, Angela
Kosson, Piotr
Florys, Katarzyna
Sączyńska, Violetta
Dębski, Janusz
Cecuda-Adamczewska, Violetta
Szewczyk, Bogusław
Zagórski-Ostoja, Włodzimierz
Grzelak, Krystyna
Powiązania:
https://bibliotekanauki.pl/articles/1039268.pdf
Data publikacji:
2014
Wydawca:
Polskie Towarzystwo Biochemiczne
Tematy:
Avian influenza
H5N1
recombinant hemagglutinin
Pichia pastoris
Opis:
The A/swan/Poland/305-135V08/2006 (H5N1-subtype) hemagglutinin (HA) gene was cloned and expressed in yeast Pichia pastoris (P. pastoris). The HA cDNA lacking the C-terminal transmembrane anchor-coding sequence was fused to an α-factor leader peptide and placed under control of the methanol-inducible P. pastoris alcohol oxidase 1 (AOX1) promoter. Two P. pastoris strains: SMD 1168 and KM 71 were used for protein expression. Recombinant HA protein was secreted into the culture medium reaching an approximately 15 mg/L (KM 71 strain). Fusion protein with a His6 tag was purified to homogeneity in one step affinity chromatography. SDS-PAGE and MS/MS analysis indicated that the protein is cleaved into HA1 and HA2 domains linked by a disulfide bond. Analysis of the N-linked glycans revealed that the overexpressed HA is fully glycosylated at the same sites as the native HA in the vaccine strain. Immunological activity of the hemagglutinin protein was tested in mice, where rHA elicited a high immune response.
Źródło:
Acta Biochimica Polonica; 2014, 61, 3; 597-602
0001-527X
Pojawia się w:
Acta Biochimica Polonica
Dostawca treści:
Biblioteka Nauki
Artykuł
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