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41.
Pseudolernentoma, a new chondracanthid genus was proposed to accommodate Pseudolernentoma brasiliensis n. g., n. sp., parasitic on the pink cusk-eel Genypterus brasiliensis Regan, from off the coast of Rio de Janeiro, Brazil. The new genus can be differentiated from the other genera of the Chondracanthidae by the presence of an inflated head with lateral expansions and anteroventral bifurcate processes on the trunk. 相似文献
42.
43.
Martha P. Brown Dimitri Toptygin K. B. Lee Theresa Animashaun R. C. Hughes Y. C. Lee Ludwig Brand 《The protein journal》1998,17(2):149-159
The plant lectin Tetracarbidium conophorum agglutinin II binds to glycoproteins and glycopeptides in a structurally specific manner [Animashaun et al., (1994) Glycoconjugate J. 11, 299–303]. We have characterized the steady-state and time-resolved fluorescence of the tryptophan residues of this lectin. The fluorescence (λex = 295 nm, λem = 350 nm) decay is complex and can be described by four decay times with the following values: τ1 = 7.4nsec, α1 = 0.22; τ2 = 2.9 nsec, α2 = 0.25; τ3 = l.0 nsec, α3 = 0.34; τ4 = 0.2 nsec, α4 = 0.18. The addition of a biantennary glycopeptide $\begin{array}{*{20}c} {Gal\beta (1 \to 4)GlcNAc\beta (1 \to 2)Man\alpha (1 \to 6)\neg } \\ {Man\beta (1 \to 4)GlcNAC\beta (1 \to 4)GlcAc\beta (1 \to )\begin{array}{*{20}c} {Glu - Nh_2 } \\ | \\ {Asn} \\ | \\ {COOH} \\ \end{array} } \\ {Gal\beta (1 \to 4)GlcNAc\beta (1 \to 2)Man\alpha (1 \to 3)} \\ \end{array} $ to the lectin results in a quench and an 8 nm blue shift of the emission spectrum. The effect is saturable, and is described by an association constant of 1.8×105 M?1. The tryptophan fluorescence of Tetracarbidium conophorum agglutinin II may therefore be utilized to characterize thermodynamically the binding interactions between this lectin and complex glycoprotein. 相似文献
44.
45.
Martha P. Brown Dimitri Toptygin K. B. Lee Theresa Animashaun R. C. Hughes Y. C. Lee Ludwig Brand 《Journal of Protein Chemistry》1998,17(2):149-159
The plant lectin Tetracarbidium conophorum agglutinin II binds to glycoproteins and glycopeptides in a structurally specific manner [Animashaun et al., (1994) Glycoconjugate J.
11, 299–303]. We have characterized the steady-state and time-resolved fluorescence of the tryptophan residues of this lectin. The fluorescence (ex = 295 nm, em = 350 nm) decay is complex and can be described by four decay times with the following values: 1 = 7.4nsec, 1 = 0.22; 2 = 2.9 nsec, 2 = 0.25; 3 = l.0 nsec, 3 = 0.34; 4 = 0.2 nsec, 4 = 0.18. The addition of a biantennary glycopeptide
to the lectin results in a quench and an 8 nm blue shift of the emission spectrum. The effect is saturable, and is described by an association constant of 1.8×105 M–1. The tryptophan fluorescence of Tetracarbidium conophorum agglutinin II may therefore be utilized to characterize thermodynamically the binding interactions between this lectin and complex glycoprotein. 相似文献
46.
Dimitri Coucouvanis 《Journal of biological inorganic chemistry》1996,1(6):594-600
Reactivity studies of clusters that contain the MFe3S4 cores (M = Mo, V) with catecholate, multicarboxylate (or DMF) ligands coordinated to the Mo (or V) atoms, and Cl ligands
coordinated to the Fe atoms have been carried out. These studies show the M/Fe/S single cubane clusters to be effective catalysts
in the reduction of nitrogenase substrates such as hydrazine, acetylene and protons to give ammonia, ethylene and dihydrogen
respectively. The same molecules do not activate or catalyze the reduction of dinitrogen. The results indicate that the observed
catalyses are occurring at the Mo (V) sites by a process that, in the case of hydrazine, involves substrate protonation prior
to reduction. The facile catalytic reduction of hydrazine by clusters that contain coordinatively saturated polycarboxylate-bound
Mo atoms is rationalized in terms of a possible protonation/proton delivery function of the coordinated polycarboxylate ligands.
The reactivity characteristics of the M/Fe/S clusters (structurally quite similar to the nitrogenase cofactor) have led to
the suggestion that the Mo (V) atoms may be involved in the reduction of hydrazine in the later stages of dinitrogen reduction.
Received and accepted: 21 August 1996 相似文献
47.
Sulaiman Al‐Zuhair Salman Ashraf Soleiman Hisaindee Naeema Al Darmaki Sinan Battah Dimitri Svistunenko Brandon Reeder Glyn Stanway Afeefa Chaudhary 《Engineering in Life Science》2017,17(2):175-185
Crude proteins and pigments were extracted from different microalgae strains, both marine and freshwater. The effectiveness of enzymatic pre‐treatment prior to protein extraction was evaluated and compared to conventional techniques, including ultrasonication and high‐pressure water extraction. Enzymatic pre‐treatment was chosen as it could be carried out at mild shear conditions and does not subject the proteins to high temperatures, as with the ultrasonication approach. Using enzymatic pre‐treatment, the extracted proteins yields of all tested microalgae strains were approximately 0.7 mg per mg of dry cell weight. These values were comparable to those achieved using a commercial lytic kit. Ultrasonication was not very effective for proteins extraction from Chlorella sp., and the extracted proteins yields did not exceed 0.4 mg per mg of dry cell weight. For other strains, similar yields were achieved by both treatment methods. The time‐course effect of enzymatic incubation on the proteins extraction efficiency was more evident using laccase compared to lysozyme, which suggested that the former enzyme has a slower rate of cell disruption. The crude extracted proteins were fractionated using an ion exchange resin and were analyzed by the electrophoresis technique. They were further tested for their antioxidant activity, the highest of which was about 60% from Nannochloropsis sp. The total phenolic contents in the selected strains were also determined, with Chlorella sp. showing the highest content reaching 17 mg/g. Lysozyme was also found to enhance the extraction of pigments, with Chlorella sp. showing the highest pigments contents of 16.02, 4.59 and 5.22 mg/g of chlorophyll a, chlorophyll b and total carotenoids, respectively. 相似文献
48.
A vector system is presented that allows generation of E. coli co-expression clones by a standardized, robust cloning procedure. The number of co-expressed proteins is not limited. Five ‘pQLink’ vectors for expression of His-tag and GST-tag fusion proteins as well as untagged proteins and for cloning by restriction enzymes or Gateway cloning were generated. The vectors allow proteins to be expressed individually; to achieve co-expression, two pQLink plasmids are combined by ligation-independent cloning. pQLink co-expression plasmids can accept an unrestricted number of genes. As an example, the co-expression of a heterotetrameric human transport protein particle (TRAPP) complex from a single plasmid, its isolation and analysis of its stoichiometry are shown. pQLink clones can be used directly for pull-down experiments if the proteins are expressed with different tags. We demonstrate pull-down experiments of human valosin-containing protein (VCP) with fragments of the autocrine motility factor receptor (AMFR). The cloning method avoids PCR or gel isolation of restriction fragments, and a single resistance marker and origin of replication are used, allowing over-expression of rare tRNAs from a second plasmid. It is expected that applications are not restricted to bacteria, but could include co-expression in other hosts such as Bacluovirus/insect cells. 相似文献
49.
Monoclonal antibody AP33 defines a broadly neutralizing epitope on the hepatitis C virus E2 envelope glycoprotein 总被引:3,自引:0,他引:3 下载免费PDF全文
Owsianka A Tarr AW Juttla VS Lavillette D Bartosch B Cosset FL Ball JK Patel AH 《Journal of virology》2005,79(17):11095-11104
Hepatitis C virus (HCV) remains a significant threat to the general health of the world's population, and there is a pressing need for the development of new treatments and preventative vaccines. Here, we describe the generation of retrovirus-based pseudoparticles (HCVpp) incorporating a panel of full-length E1E2 clones representative of the major genotypes 1 through 6, and their application to assess the reactivity and neutralizing capability of antisera and monoclonal antibodies raised against portions of the HCV E2 envelope protein. Rabbit antisera raised against either the first hypervariable region or ectodomain of E2 showed limited and strain specific neutralization. By contrast, the monoclonal antibody (MAb) AP33 demonstrated potent neutralization of infectivity against HCVpp carrying E1E2 representative of all genotypes tested. The concentration of AP33 required to achieve 50% inhibition of infection by HCVpp of diverse genotypes ranged from 0.6 to 32 mug/ml. The epitope recognized by MAb AP33 is linear and highly conserved across different genotypes of HCV. Thus, identification of a broadly neutralizing antibody that recognizes a linear epitope is likely to be of significant benefit to future vaccine and therapeutic antibody development. 相似文献
50.
Relationship between SU subdomains that regulate the receptor-mediated transition from the native (fusion-inhibited) to the fusion-active conformation of the murine leukemia virus glycoprotein 下载免费PDF全文
Envelope glycoproteins (Env) of retroviruses are trimers of SU (surface) and TM (transmembrane) heterodimers and are expressed on virions in fusion-competent forms that are likely to be metastable. Activation of the viral receptor-binding domain (RBD) via its interaction with a cell surface receptor is thought to initiate a cascade of events that lead to refolding of the Env glycoprotein into its stable fusion-active conformation. While the fusion-active conformation of the TM subunit has been described in detail for several retroviruses, little is known about the fusion-competent structure of the retroviral glycoproteins or the molecular events that mediate the transition between the two conformations. By characterizing Env chimeras between the ecotropic and amphotropic murine leukemia virus (MLV) SUs as well as a set of point mutants, we show that alterations of the conformation of the SU glycoprotein strongly elevate Env fusogenicity by disrupting the stability of the Env complex. Compensatory mutations that restored both Env stability and fusion control were also identified, allowing definition of interactions within the Env complex that maintain the stability of the native Env complex. We show that, in the receptor-unbound form, structural interactions between the N terminus of the viral RBD (NTR domain), the proline-rich region (PRR), and the distal part of the C-terminal domain of the SU subunit maintain a conformation of the glycoprotein that is fusion inhibitory. Additionally, we identified mutations that disrupt this fusion-inhibitory conformation and allow fusion activation in the absence of viral receptors, provided that receptor-activated RBD fragments are added in trans during infection. Other mutations were identified that allow fusion activation in the absence of receptors for both the viral glycoprotein and the trans-acting RBD. Finally, we found mutations of the SU that bypass in cis the requirement for the NTR domain in fusion activation. All these different mutations call for a critical role of the PRR in mediating conformational changes of the Env glycoprotein during fusion activation. Our results suggest a model of MLV Env fusion activation in which unlocking of the fusion-inhibitory conformation is initiated by receptor binding of the viral RBD, which, upon disruption of the PRR, allows the NTR domain to promote further events in Env fusion activation. This involves a second type of interaction, in cis or in trans, between the receptor-activated RBD and a median segment of the freed C-terminal domain. 相似文献