Lectins with a β-propeller fold bind glycans on the cell surface through multivalent binding sites and appropriate directionality. These proteins are formed by repeats of short domains, raising questions about evolutionary duplication. However, these repeats are difficult to detect in translated genomes and seldom correctly annotated in sequence databases. To address these issues, we defined the blade signature of the five types of β-propellers using 3D-structural data. With these templates, we predicted 3,887 β-propeller lectins in 1,889 species and organized this information in a searchable online database. The data reveal a widespread distribution of β-propeller lectins across species. Prediction also emphasizes multiple architectures and led to the discovery of a β-propeller assembly scenario. This was confirmed by producing and characterizing a predicted protein coded in the genome of Kordia zhangzhouensis. The crystal structure uncovers an intermediate in the evolution of β-propeller assembly and demonstrates the power of our tools.
- Klíčová slova
- carbohydrate binding protein, lectins, oligomerization, β-propeller,
- MeSH
- Archaea chemie MeSH
- Bacteria chemie MeSH
- databáze proteinů MeSH
- Eukaryota chemie MeSH
- genom bakteriální MeSH
- lektiny chemie MeSH
- molekulární modely MeSH
- multimerizace proteinu MeSH
- proteom MeSH
- sbalování proteinů MeSH
- sekundární struktura proteinů MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- vazebná místa MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- lektiny MeSH
- proteom MeSH
Polarization microscopy has been used to study the internal structures of microbial cells and in terms of the birefringence of these structures and its possible relation to the cell function and composition. Cyanobacteria of the genus Phormidium were found to contain no anisotropic structures, while other microorganisms were found to contain them, albeit to a different extent, size, and number. The flagellate Euglena was found to contain two large anisotropic bodies, whereas the flagellate of the genus Phacus belonging to the same systematic group Euglenales was observed to contain only one large anisotropic body (storage substances--paramylon). On the other hand, green algae of the genus Scenedesmus, whose cells form four--celled coenobia, contained clusters of small anisotropic granules composed also of storage substances (volutin). Minute anisotropic granules (storage substances) in two smaller clusters were found also in diatoms of the genus Navicula, whereas the green alga of the genus Mougeotia was revealed to contain, in addition to minute anisotropic granules (storage substances) occurring in low numbers in the cytoplasm, also a strongly birefringent cell wall (shape birefringence). Cells of the amoeba of the genus Naegleria and heliozoans of the genus Heterophrys were observed to contain only isolated tiny anisotropic granules (storage substances).
BACKGROUND: RanBPM (Ran-binding protein in the microtubule-organizing centre) was originally reported as a centrosome-associated protein in human cells. However, RanBPM protein containing highly conserved SPRY, LisH, CTLH and CRA domains is currently considered as a scaffolding protein with multiple cellular functions. A plant homologue of RanBPM has not yet been characterized. RESULTS: Based on sequence similarity, we identified a homologue of the human RanBPM in Arabidopsis thaliana. AtRanBPM protein has highly conserved SPRY, LisH, CTLH and CRA domains. Cell fractionation showed that endogenous AtRanBPM or expressed GFP-AtRanBPM are mainly cytoplasmic proteins with only a minor portion detectable in microsomal fractions. AtRanBPM was identified predominantly in the form of soluble cytoplasmic complexes ~230-500 kDa in size. Immunopurification of AtRanBPM followed by mass spectrometric analysis identified proteins containing LisH and CRA domains; LisH, CRA, RING-U-box domains and a transducin/WD40 repeats in a complex with AtRanBPM. Homologues of identified proteins are known to be components of the C-terminal to the LisH motif (CTLH) complexes in humans and budding yeast. Microscopic analysis of GFP-AtRanBPM in vivo and immunofluorescence localization of endogenous AtRanBPM protein in cultured cells and seedlings of Arabidopsis showed mainly cytoplasmic and nuclear localization. Absence of colocalization with γ-tubulin was consistent with the biochemical data and suggests another than a centrosomal role of the AtRanBPM protein. CONCLUSION: We showed that as yet uncharacterized Arabidopsis RanBPM protein physically interacts with LisH-CTLH domain-containing proteins. The newly identified high molecular weight cytoplasmic protein complexes of AtRanBPM showed homology with CTLH types of complexes described in mammals and budding yeast. Although the exact functions of the CTLH complexes in scaffolding of protein degradation, in protein interactions and in signalling from the periphery to the cell centre are not yet fully understood, structural conservation of the complexes across eukaryotes suggests their important biological role.
- MeSH
- adaptorové proteiny signální transdukční genetika metabolismus MeSH
- Arabidopsis chemie genetika metabolismus MeSH
- cytoskeletální proteiny genetika metabolismus MeSH
- Eukaryota chemie klasifikace genetika MeSH
- jaderné proteiny genetika metabolismus MeSH
- konzervovaná sekvence MeSH
- lidé MeSH
- molekulární evoluce * MeSH
- molekulární sekvence - údaje MeSH
- proteiny huseníčku chemie genetika metabolismus MeSH
- rostliny chemie klasifikace genetika MeSH
- sekvence aminokyselin MeSH
- sekvenční homologie aminokyselin MeSH
- terciární struktura proteinů MeSH
- vazba proteinů MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- adaptorové proteiny signální transdukční MeSH
- cytoskeletální proteiny MeSH
- jaderné proteiny MeSH
- proteiny huseníčku MeSH
- Ran binding protein 9 MeSH Prohlížeč
A new extraction technique based on the off-line combination of pressurized-liquid with solid-phase extraction (PLE-SPE) is described. The method was used for the extraction of bioactive phenolic acids (protocatechuic, p-hydroxybenzoic, 2,3-dihydroxybenzoic, chlorogenic, vanillic, caffeic, p-coumaric, salicylic acid), cinnamic acid and hydroxybenzaldehydes (p-hydroxybenzaldehyde, 3,4-dihydroxybenzaldehyde, vanillin) from in vitro culture of two freshwater algae (Anabaena doliolum and Spongiochloris spongiosa) and from food products of marine macroalgae Porphyra tenera (nori) and Undaria pinnatifida (wakame). For the identification and quantification of the compounds the molecular ions [M-H](-) and specific fragments were analyzed by quadrupole mass spectrometry analyzer connected on-line with a reversed-phase HPLC system. Our analysis showed that the freshwater algae and marine algal products contained submicrogram or microgram level of above-mentioned phenols per gram of lyophilized sample. In addition, the total phenol content (Folin-Ciocalteu assay) and antioxidant activity (TEAC assay, Trolox equivalent antioxidant capacity assay) of the PLE-SPE extracts were determined and discussed.
- MeSH
- antioxidancia analýza chemie MeSH
- biologie sladkých vod metody MeSH
- cinnamáty analýza chemie MeSH
- Eukaryota chemie MeSH
- extrakce na pevné fázi metody MeSH
- fenoly analýza chemie MeSH
- kyselina benzoová analýza chemie MeSH
- mořská voda chemie MeSH
- on-line systémy * MeSH
- referenční standardy MeSH
- vysokoúčinná kapalinová chromatografie metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antioxidancia MeSH
- cinnamáty MeSH
- cinnamic acid MeSH Prohlížeč
- fenoly MeSH
- kyselina benzoová MeSH
An important component of the photosynthetic apparatus is a light-harvesting system that captures light energy and transfers it efficiently to the reaction center. Depending on environmental conditions, photosynthetic antennae have adopted various strategies for this function. Peridinin-chlorophyll-a protein (PCP) represents a unique situation because, unlike other antenna systems which have a preponderance of chlorophyll, it has the carotenoid, peridinin, as its major pigment. The key structural feature of peridinin is a conjugated carbonyl group. Owing to the presence of this group, an intramolecular charge-transfer excited state is formed in peridinin which exhibits different excited state spectra and dynamics depending on the polarity of the environment. The charge-transfer state also facilitates energy transfer between peridinin and chlorophyll-a in PCP. This review summarizes results of spectroscopic investigations of PCP in the past few years, emphasizing the specific light-harvesting strategy developed by marine photosynthetic organisms utilizing carbonyl-containing carotenoids in their antenna complexes.
- MeSH
- chlorofyl chemie MeSH
- Eukaryota chemie fyziologie MeSH
- fotosyntéza fyziologie MeSH
- karotenoidy chemie MeSH
- nukleární magnetická rezonance biomolekulární MeSH
- přenos energie MeSH
- světlosběrné proteinové komplexy chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- chlorofyl MeSH
- karotenoidy MeSH
- peridinin MeSH Prohlížeč
- světlosběrné proteinové komplexy MeSH
An on-line coupled capillary isotachophoresis--capillary zone electrophoresis (cITP-CZE) method for the determination of domoic acid in shellfish and algae is described. The optimised cITP-CZE electrolyte system was 10 mM HCl + 20 mM beta-alanine (BALA) + 0.05% hydroxyethylcellulose (leading electrolyte), 5 mM caproic acid (terminating electrolyte) and 20 mM caproic acid + 20 mM BALA + 0.1% HPMC (background electrolyte). A clear separation of the domoic acid from the other components of methanolic sample extract was achieved within 25 min. Method characteristics, i.e., linearity (0-200 microg/l), accuracy (recovery 101+/-3%), intra-assay repeatability (2.4%) and detection limit (1.5 microg/l) were determined. Speed of analysis, low laboriousness, high sensitivity and low running cost are the typical attributes of the cITP-CZE method. Developed method was successfully applied to analysis of shellfish samples and food supplements containing algae extract.
- MeSH
- elektroforéza kapilární metody MeSH
- elektroforéza metody MeSH
- Eukaryota chemie MeSH
- kalibrace MeSH
- kyselina kainová analogy a deriváty analýza MeSH
- měkkýši, korýši analýza MeSH
- referenční standardy MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- domoic acid MeSH Prohlížeč
- kyselina kainová MeSH
Phylogeny of seven groups of metazoan parasitic groups is reviewed, based on both morphological and molecular data. The Myxozoa (=Malacosporea + Myxosporea) are most probably related to the egg-parasitic cnidarian Polypodium (Hydrozoa?: Polypodiozoa); the other phylogenetic hypotheses are discussed and the possible non-monophyly of the Cnidaria (with the Polypodiozoa-Myxozoa clade closest to the Triploblastica) is suggested. The Mesozoa is a monophyletic group, possibly closely related to the (monophyletic) Acoelomorpha; whether the Acoelomorpha and Mesozoa represent the basalmost triploblast clade(s) or a derived platyhelminth subclade may depend on rooting the tree of the Triploblastica. Position of the monophyletic Neodermata (=Trematoda + Cercomeromorpha) within the rhabditophoran flatworms is discussed, with two major alternative hypotheses about the neodermatan sister-group relationships (viz., the "neoophoran" and "revertospermatan"). The Myzostomida are not annelids but belong among the Platyzoa, possibly to the clade of animals with anterior sperm flagella (=Prosomastigozoa). The Acanthocephala represent derived syndermates ("rotifers"), possibly related to Seison (the name Pararotatoria comb. n. is proposed for Seisonida + Acanthocephala). The crustacean origin of the Pentastomida based on spermatological and molecular evidence (Pentastomida + Branchiura = Ichthyostraca) is confronted with palaeontological views favouring the pre-arthropod derivation of the pentastomids. Phylogenetic position of the nematodes within the Ecdysozoa and evolution of nematode parasitism are discussed, and the lack of relevant information about the enigmatic ectoproctan parasite Buddenbrockia is emphasised.
- MeSH
- bezobratlí klasifikace genetika MeSH
- cizopasní červi chemie klasifikace genetika MeSH
- DNA genetika MeSH
- Eukaryota chemie klasifikace genetika MeSH
- fylogeneze * MeSH
- molekulární evoluce * MeSH
- ribozomální DNA genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- DNA MeSH
- ribozomální DNA MeSH
A histochemical study using lectin methods was performed on myxosporean parasites from vastly different fish hosts from marine and fresh waters. Six biotinylated lectins were used (WGA, SBA, BS-I, Con-A, UEA-I and SNA). The binding pattern of Con-A and WGA revealed the presence of mannose and/or glucose, and N-acetyl-D-glucosamine respectively, in polar capsules and valves of most of the myxosporea assayed. Thus, chitin may be present in polar capsules and/or valves of myxosporean spores. The BS-I binding pattern showed the presence of alpha-D-galactose and/or N-acetyl-D-galactosamine residues in polar capsules of Kudoa sp., Zschokkella mugilis Sitjà-Bobadilla et Alvarez-Pellitero, 1993 and Leptotheca sp., and in the valves of the latter. Scarce amounts of N-acetyl-D-galactosamine and/or alpha-D-galactose were demonstrated by SBA binding in Sphaerospora dicentrarchi Sitjà-Bobadilla et Alvarez-Pellitero 1992, Leptotheca sp. and Kudoa sp. valves, and in Leptotheca sp. polar capsules. The UEA-I staining indicated the absence of alpha-L-fucose in all the myxosporea assayed except in Leptotheca sp. N-acetylneuraminic acid was detected with SNA in the polar capsules and sporoplasms of Polysporoplasma sparis Sitjà-Bobadilla et Alvarez-Pellitero, 1995 and in the polar capsules and valves of Kudoa sp. These results indicate that, although Myxosporea may have conserved carbohydrate structures, some of them can show significantly different binding patterns, which may be useful in diagnostic and functional studies.
- MeSH
- biotinylace MeSH
- Eukaryota chemie metabolismus MeSH
- histocytochemie MeSH
- lektiny metabolismus MeSH
- metabolismus sacharidů MeSH
- molekulární sekvence - údaje MeSH
- nemoci ryb parazitologie MeSH
- Perciformes parazitologie MeSH
- protozoální infekce zvířat parazitologie MeSH
- sacharidové sekvence MeSH
- sacharidy analýza chemie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- lektiny MeSH
- sacharidy MeSH
