This review article provides a wide overview of important developments and applications of capillary electromigration methods in the area of peptide mapping of proteins in the period 1997-mid-2022, including review articles on this topic. It deals with all major aspects of peptide mapping by capillary electromigration methods: i) precleavage sample preparation involving purification, preconcentration, denaturation, reduction and alkylation of protein(s) to be analyzed, ii) generation of peptide fragments by off-line or on-line enzymatic and/or chemical cleavage of protein(s), iii) postcleavage preparation of the generated peptide mixture for capillary electromigration separation, iv) separation of the complex peptide mixtures by one-, two- and multidimensional capillary electromigration methods coupled with mass spectrometry detection, and v) a large application of peptide mapping for variable purposes, such as qualitative analysis of monoclonal antibodies and other protein biopharmaceuticals, monitoring of posttranslational modifications, determination of primary structure and investigation of function of proteins in biochemical and clinical research, characterization of proteins of variable origin as well as for protein and peptide identification in proteomic and peptidomic studies.
- Klíčová slova
- capillary electrophoresis, peptide fingerprint, peptides, proteins, review,
- MeSH
- peptidové mapování MeSH
- peptidy * MeSH
- proteomika * MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- peptidy * MeSH
This paper provides a detailed overview of relevant developments in separation and analysis of proteins by capillary electromigration (CE) methods in the period 2017-mid 2021. The presented topics embrace sample preparation, suppression of protein sorption, control of electroosmotic flow, separations of proteins by the particular CE methods and protein identification and quantification by various detection techniques. To illustrate the wide usability of CE methods for protein analysis, the paper shows several novel applications of CE methods, such as quality control and characterization of protein biopharmaceuticals, determination of proteins in complex biomatrices, investigation of posttranslational modification, peptide mapping, interactions of proteins with other (bio)molecules and determination of their physicochemical and biochemical characteristics.
- Klíčová slova
- Affinity electrophoresis, Capillary electrochromatography, Capillary electrophoresis, Isoelectric focusing, Proteins, Review,
- MeSH
- elektroforéza kapilární * metody MeSH
- elektroosmóza MeSH
- peptidové mapování MeSH
- peptidy * chemie MeSH
- proteiny analýza MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- peptidy * MeSH
- proteiny MeSH
The review provides a comprehensive overview of developments and applications of high performance capillary and microchip electroseparation methods (zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography) for analysis, microscale isolation, and physicochemical characterization of peptides from 2019 up to approximately the middle of 2021. Advances in the investigation of electromigration properties of peptides and in the methodology of their analysis, such as sample preparation, sorption suppression, EOF control, and detection, are presented. New developments in the individual CE and CEC methods are demonstrated and several types of their applications are shown. They include qualitative and quantitative analysis, determination in complex biomatrices, monitoring of chemical and enzymatic reactions and physicochemical changes, amino acid, sequence, and chiral analyses, and peptide mapping of proteins. In addition, micropreparative separations and determination of significant physicochemical parameters of peptides by CE and CEC methods are described.
- Klíčová slova
- Capillary electrochromatography, Capillary electrophoresis, Peptides, Proteins, Review,
- MeSH
- elektroforéza kapilární * MeSH
- isoelektrická fokusace MeSH
- peptidové mapování MeSH
- peptidy * izolace a purifikace MeSH
- proteiny analýza MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- peptidy * MeSH
- proteiny MeSH
The review presents a comprehensive survey of recent developments and applications of high performance capillary and microchip electroseparation methods (zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography) for analysis, micropreparation, and physicochemical and biochemical characterization of peptides since 2017 up to about the middle of 2019. Progress in the study of electromigration properties of peptides and in the methodology of their analysis (sample preseparation, preconcentration and derivatization, adsorption suppression, EOF control, and detection) are described. Advances in CE and CEC methods are demonstrated and their applications in the following areas are presented: qualitative and quantitative analysis, determination in complex (bio)matrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid, sequence and chiral analysis, and peptide mapping of proteins. In addition, micropreparative separations and determinations of important physicochemical characteristics of peptides by CE and CEC methods are reported.
- Klíčová slova
- Capillary electrochromatography, Capillary electrophoresis, Peptides, Proteins, Review,
- MeSH
- elektroforéza kapilární * MeSH
- kapilární elektrochromatografie * MeSH
- peptidové mapování MeSH
- peptidy analýza chemie izolace a purifikace MeSH
- proteiny analýza chemie izolace a purifikace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- peptidy MeSH
- proteiny MeSH
CD molecules are surface molecules expressed on cells of the immune system that play key roles in immune cell-cell communication and sensing the microenvironment. These molecules are essential markers for the identification and isolation of leukocytes and lymphocyte subsets. Here, we present the results of the first phase of the CD Maps study, mapping the expression of CD1-CD100 (n = 110) on 47 immune cell subsets from blood, thymus, and tonsil using an eight-color standardized EuroFlow approach and quantification of expression. The resulting dataset included median antibody binding capacities (ABCs) and percentage of positivity for all markers on all subsets and was developed into an interactive CD Maps web resource. Using the resource, we examined differentially expressed proteins between granulocyte, monocyte, and dendritic cell subsets, and profiled dynamic expression of markers during thymocyte differentiation, T-cell maturation, and between functionally distinct B-cell subset clusters. The CD Maps resource will serve as a benchmark of antibody reactivities ensuring improved reproducibility of flow cytometry-based research. Moreover, it will provide a full picture of the surfaceome of human immune cells and serves as a useful platform to increase our understanding of leukocyte biology, as well as to facilitate the identification of new biomarkers and therapeutic targets of immunological and hematological diseases.
- Klíčová slova
- B-cell, CD marker, T-cell, expression profiling, flow cytometry, lymphocyte, monocyte, surfaceome,
- MeSH
- B-lymfocyty imunologie metabolismus MeSH
- CD antigeny biosyntéza MeSH
- datové soubory jako téma MeSH
- dendritické buňky imunologie metabolismus MeSH
- dítě MeSH
- dospělí MeSH
- granulocyty imunologie metabolismus MeSH
- imunofenotypizace MeSH
- internet MeSH
- leukocyty imunologie metabolismus MeSH
- lidé MeSH
- lymfopoéza MeSH
- monocyty imunologie metabolismus MeSH
- peptidové mapování MeSH
- podskupiny lymfocytů imunologie metabolismus MeSH
- předškolní dítě MeSH
- průtoková cytometrie MeSH
- reprodukovatelnost výsledků MeSH
- separace buněk MeSH
- T-lymfocyty imunologie metabolismus MeSH
- Check Tag
- dítě MeSH
- dospělí MeSH
- lidé MeSH
- předškolní dítě MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- CD antigeny MeSH
The present study is focused on the characterization of yacon [Smallanthus sonchifolius (Poepp. et Endl.) H. Robinson] accessions from different geographic origins (Bolivia, Ecuador, and Peru) by iPBS markers and metabolomic fingerprinting. The results showed that the number of amplified polymorphic fragment levels ranged from 20 up to 27 with a level of polymorphism ranging from 80 to 100%. Five of the iPBS primers used in this study provided no specific banding pattern able to discriminate between the different yacon accessions. However, two iPBS primer pairs were able to separate Peru accessions from those of Ecuador and Bolivia. The UPLC-HRMS/MS-based metabolomic fingerprinting showed highly similar metabolomic fingerprints characterized by the accumulation of high quantities of sesquiterpene lactones and diterpenes, but no apparent geographic clustering. The present study demonstrates that yacon accessions from different geographical origins maintained ex situ (in the Czech Republic) present a rather low chemical and genetic diversity.
- Klíčová slova
- Genetic diversity, Metabolomic fingerprints, Retrotransposons, Smallanthus sonchifolius, iPBS,
- MeSH
- antioxidancia chemie MeSH
- Asteraceae chemie genetika MeSH
- diterpeny chemie MeSH
- genetická variace MeSH
- glykosylace MeSH
- hmotnostní spektrometrie MeSH
- kořeny rostlin chemie MeSH
- laktony chemie MeSH
- metabolomika MeSH
- multivariační analýza MeSH
- peptidové mapování MeSH
- retroelementy MeSH
- rostlinné extrakty chemie MeSH
- seskviterpeny chemie MeSH
- shluková analýza MeSH
- zeměpis MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Bolívie MeSH
- Česká republika MeSH
- Ekvádor MeSH
- Peru MeSH
- Názvy látek
- antioxidancia MeSH
- diterpeny MeSH
- laktony MeSH
- retroelementy MeSH
- rostlinné extrakty MeSH
- seskviterpeny MeSH
We studied the taxonomic position of six phenetically related strains of the genus Acinetobacter, which were recovered from hospital sewage in China and showed different patterns of resistance to clinically important antibiotics. Whole-genome sequencing of these strains and genus-wide phylogeny reconstruction based on a set of 107 Acinetobacter core genes indicated that they formed a separate and internally cohesive clade within the genus. The average nucleotide identity based on BLAST and digital DNA-DNA hybridization values between the six new genomes were 97.25-98.67% and 79.2-89.3%, respectively, whereas those between them and the genomes of the known species were ≤78.57% and ≤28.5%, respectively. The distinctness of the strains at the species level was also supported by the results of the cluster analysis of the whole-cell protein fingerprints generated by MALDI-TOF MS. Moreover, the strains displayed a catabolically unique profile and could be differentiated from the phylogenetically closest species at least by their inability to grow on d,l-lactate. A total of 18 different genes were found in the six genome sequences which encode resistance to seven classes of antimicrobial agents, including clinically important carbapenems, oxyimino-cephalosporins, or aminoglycosides. These genes occurred in five different combinations, with three to 10 different genes per strain. We conclude that the six strains represent a novel Acinetobacter species, for which we propose the name Acinetobacter cumulans sp. nov. to reflect its ability to acquire and cumulate diverse resistance determinants. The type strain is WCHAc060092T (ANC 5797T=CCTCC AB 2018119T=GDMCC 1.1380T=KCTC 62576T).
- Klíčová slova
- Carbon source assimilation, Core genome phylogeny, MALDI-TOF MS, Whole genome sequence,
- MeSH
- Acinetobacter chemie klasifikace účinky léků genetika MeSH
- antibakteriální látky farmakologie MeSH
- bakteriální geny * MeSH
- DNA bakterií genetika MeSH
- fylogeneze MeSH
- genom bakteriální genetika MeSH
- hybridizace nukleových kyselin MeSH
- mnohočetná bakteriální léková rezistence genetika MeSH
- nemocnice * MeSH
- odpadní vody mikrobiologie MeSH
- peptidové mapování MeSH
- RNA ribozomální 16S genetika MeSH
- sekvenční analýza DNA MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Čína MeSH
- Názvy látek
- antibakteriální látky MeSH
- DNA bakterií MeSH
- odpadní vody MeSH
- RNA ribozomální 16S MeSH
The objective of this study was to investigate the patient-reported outcomes (PROs) and matrix metalloproteinase (MMP) derived extracellular matrix (ECM) biomarkers in non-radiographic (nr)-axial spondyloarthritis (axSpA) and radiographic (r)-axSpA after exercise intervention. Forty-six axSpA patients with stable disease and treatment underwent 24 weeks long exercise intervention. The clinical and laboratory assessments were performed at baseline and at follow-up. The PROs included evaluation of patient's global disease activity (PGDA), disease activity (DA7), pain (PAIN7) and fatigue during last week and quality of life questionnaires. ELISAs for MMP-degraded collagen type II, C-reactive protein (CRPM) and citrullinated vimentin were used. The data of 23 r-axSpA and 19 nr-axSpA were analysed. The PDGA was similar for nr-axSpA (35.2 ± 18.9) and r-axSpA (33.4 ± 22.3) at baseline, improved significantly after intervention (p < 0.01) and the change of PDGA was almost identical for nr-axSpA (- 10.0 ± 15.4) and r-axSpA (- 9.8 ± 11.9). Evaluations of DA7 and PAIN7 were significantly improved only in nr-axSpA (3.5 ± 2.3 and 34.7 ± 25.6 at baseline vs. 2.1 ± 1.9 and 21.0 ± 20.5, respectively, p < 0.01). The decline of DA7 and PAIN7 was more profound, but not significantly in nr-axSpA than in r-axSpA (- 1.4 ± 1.6 and - 13.7 ± 17.4 vs. - 0.5 ± 3.1 and - 3.7 ± 3.3, respectively). The quality of life was not changed. At baseline, increased levels of CRPM were found in r-axSpA (14.85 ± 4.10) compared to nr-axSpA (11.83 ± 3.20), p < 0.05, but all three biomarkers were not influenced by exercise therapy. We found that exercise therapy mainly in the nr-axSpA improves PROs, but not ECM turnover biomarkers. This indicates that exercise therapy is important for patients' health but does not affect ECM turnover.
- Klíčová slova
- axial spondyloarthritis, exercise therapy, extracellular matrix, matrix metalloproteinase,
- MeSH
- biologické markery analýza MeSH
- C-reaktivní protein analýza MeSH
- dospělí MeSH
- hodnocení výsledků péče pacientem * MeSH
- kvalita života MeSH
- lidé MeSH
- matrixové metaloproteinasy analýza MeSH
- peptidové mapování MeSH
- spondylartritida rehabilitace MeSH
- terapie cvičením * MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biologické markery MeSH
- C-reaktivní protein MeSH
- matrixové metaloproteinasy MeSH
The F-ATPases (also called the F1 Fo -ATPases or ATP synthases) are multi-subunit membrane-bound molecular machines that produce ATP in bacteria and in eukaryotic mitochondria and chloroplasts. The structures and enzymic mechanisms of their F1 -catalytic domains are highly conserved in all species investigated hitherto. However, there is evidence that the F-ATPases from the group of protozoa known as Euglenozoa have novel features. Therefore, we have isolated pure and active F1 -ATPase from the euglenozoan parasite, Trypanosoma brucei, and characterized it. All of the usual eukaryotic subunits (α, β, γ, δ, and ε) were present in the enzyme, and, in addition, two unique features were detected. First, each of the three α-subunits in the F1 -domain has been cleaved by proteolysis in vivo at two sites eight residues apart, producing two assembled fragments. Second, the T. brucei F1 -ATPase has an additional subunit, called p18, present in three copies per complex. Suppression of expression of p18 affected in vitro growth of both the insect and infectious mammalian forms of T. brucei. It also reduced the levels of monomeric and multimeric F-ATPase complexes and diminished the in vivo hydrolytic activity of the enzyme significantly. These observations imply that p18 plays a role in the assembly of the F1 domain. These unique features of the F1 -ATPase extend the list of special characteristics of the F-ATPase from T. brucei, and also, demonstrate that the architecture of the F1 -ATPase complex is not strictly conserved in eukaryotes.
- Klíčová slova
- Trypanosoma brucei, ATP synthase, F1-domain, p18-subunit, proteolysis of α-subunit, subunit composition,
- MeSH
- adenosintrifosfát metabolismus MeSH
- hydrolýza MeSH
- kinetika MeSH
- konformace proteinů MeSH
- konzervovaná sekvence MeSH
- membránový potenciál mitochondrií MeSH
- molekulární modely * MeSH
- multimerizace proteinu MeSH
- peptidové mapování MeSH
- podjednotky proteinů antagonisté a inhibitory genetika izolace a purifikace metabolismus MeSH
- proteolýza MeSH
- protonové ATPasy antagonisté a inhibitory genetika izolace a purifikace metabolismus MeSH
- protozoální proteiny antagonisté a inhibitory genetika izolace a purifikace metabolismus MeSH
- RNA interference MeSH
- sekvence aminokyselin MeSH
- sekvenční homologie aminokyselin MeSH
- sekvenční seřazení MeSH
- stabilita enzymů MeSH
- Trypanosoma brucei brucei enzymologie růst a vývoj MeSH
- výpočetní biologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- adenosintrifosfát MeSH
- podjednotky proteinů MeSH
- protonové ATPasy MeSH
- protozoální proteiny MeSH
The review brings a comprehensive overview of recent developments and applications of high performance capillary and microchip electroseparation methods (zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography) to analysis, microscale isolation, purification, and physicochemical and biochemical characterization of peptides in the years 2015, 2016, and ca. up to the middle of 2017. Advances in the investigation of electromigration properties of peptides and in the methodology of their analysis (sample preseparation, preconcentration and derivatization, adsorption suppression and EOF control, and detection) are described. New developments in particular CE and CEC methods are presented and several types of their applications to peptide analysis are reported: qualitative and quantitative analysis, determination in complex (bio)matrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid, sequence and chiral analysis, and peptide mapping of proteins. Some micropreparative peptide separations are shown and capabilities of CE and CEC methods to provide important physicochemical characteristics of peptides are demonstrated.
- Klíčová slova
- Capillary electrochromatography, Capillary electrophoresis, Peptides, Proteins, Review,
- MeSH
- aminokyseliny chemie MeSH
- chemická frakcionace MeSH
- elektroforéza kapilární metody MeSH
- lidé MeSH
- mikročipová analýza metody MeSH
- peptidové mapování MeSH
- peptidy analýza izolace a purifikace MeSH
- stereoizomerie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- aminokyseliny MeSH
- peptidy MeSH
