scholarly journals Characterization of a minus end-directed kinesin-like motor protein from cultured mammalian cells.

1995 ◽  
Vol 129 (4) ◽  
pp. 1049-1059 ◽  
Author(s):  
R Kuriyama ◽  
M Kofron ◽  
R Essner ◽  
T Kato ◽  
S Dragas-Granoic ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Amino Acid ◽  
Mammalian Cells ◽  
Amino Acid Residues ◽  
Central Stalk ◽  
Vitro Analysis ◽  
Kinesin Superfamily ◽  
In Vitro Analysis

Using the CHO2 monoclonal antibody raised against CHO spindles (Sellitto, C., M. Kimble, and R. Kuriyama. 1992. Cell Motil. Cytoskeleton. 22:7-24) we identified a 66-kD protein located at the interphase centrosome and mitotic spindle. Isolated cDNAs for the antigen encode a 622-amino acid polypeptide. Sequence analysis revealed the presence of 340-amino acid residues in the COOH terminus, which is homologous to the motor domain conserved among other members of the kinesin superfamily. The protein is composed of a central alpha-helical portion with globular domains at both NH2 and COOH termini, and the epitope to the monoclonal antibody resides in the central alpha-helical stalk. A series of deletion constructs were created for in vitro analysis of microtubule interactions. While the microtubule binding and bundling activities require both the presence of the COOH terminus and the alpha-helical domain, the NH2-terminal half of the antigen lacked the ability to interact with microtubules. The full-length as well as deleted proteins consisting of the COOH-terminal motor and the central alpha-helical stalk supported microtubule gliding, with velocity ranging from 1.0 to 8.4 microns/minute. The speed of microtubule movement decreased with decreasing lengths of the central stalk attached to the COOH-terminal motor. The microtubules moved with their plus end leading, indicating that the antigen is a minus end-directed motor. The CHO2 sequence shows 86% identify to HSET, a gene located at the centromeric end of the human MHC region in chromosome 6 (Ando, A., Y. Y. Kikuti, H. Kawata, N. Okamoto, T. Imai, T. Eki, K. Yokoyama, E. Soeda, T. Ikemura, K. Abe, and H. Inoko. 1994. Immunogenetics. 39:194-200), indicating that HSET might represent a human homologue of the CHO2 antigen.

scholarly journals Anti-Inflammatory Formulation and Characterization of an Herbal Formulation

2021 ◽  
pp. 51-57
Author(s):  
Snigdha Pattnaik ◽  
Laxmidhar Moharana
Keyword(s):  
Pain Reduction ◽  
Inflammatory Activity ◽  
Topical Gel ◽  
Herbal Formulation ◽  
Characterization Studies ◽  
Anti Inflammatory ◽  
Vitro Analysis ◽  
In Vitro Analysis

Anti-inflammatory refers to group of medication that is utilized for curing pain and inflammation. Many synthetic products are used as anti-inflammatory agents but the effects caused by them is not satisfactory. Cordis obliqua is an anti-inflammatory herb that is enormously involved in pain reduction. The present paper aims to develop a topical gel formulation comprising herbs i.e. Cordis obliqua and curcumin with potent anti-inflammatory activity. The gel formulation was then subjected to characterization studies which involve pH, viscosity, excrudability, spreadability and in vitro analysis. pH of gel formulation was found to be 7.2±1.01 which is almost equivalent to the skin pH. In viscosity test, it was observed that viscosity of gel formulation was found to be effective. Moreover, spreadability co-efficient of the formulation satisfactory which indicates that sanitizer gel easily adhers on the skin and covers the bacteria residing on the skin, if any.

scholarly journals In vitro Characterization of Fitness and Convalescent Antibody Neutralization of SARS-CoV-2 Cluster 5 Variant Emerging in Mink at Danish Farms

2021 ◽  
Vol 12 ◽  
Author(s):  
Ria Lassaunière ◽  
Jannik Fonager ◽  
Morten Rasmussen ◽  
Anders Frische ◽  
Charlotta Polacek ◽  
...  
Keyword(s):  
Amino Acid ◽  
Reference Strain ◽  
Rhesus Macaques ◽  
Amino Acid Residues ◽  
Susceptible Animal ◽  
In Vitro Characterization ◽  
Cynomolgus Macaques ◽  
Animal Hosts

In addition to humans, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can transmit to animals that include hamsters, cats, dogs, mink, ferrets, tigers, lions, cynomolgus macaques, rhesus macaques, and treeshrew. Among these, mink are particularly susceptible. Indeed, 10 countries in Europe and North America reported SARS-CoV-2 infection among mink on fur farms. In Denmark, SARS-CoV-2 spread rapidly among mink farms and spilled-over back into humans, acquiring mutations/deletions with unknown consequences for virulence and antigenicity. Here we describe a mink-associated SARS-CoV-2 variant (Cluster 5) characterized by 11 amino acid substitutions and four amino acid deletions relative to Wuhan-Hu-1. Temporal virus titration, together with genomic and subgenomic viral RNA quantitation, demonstrated a modest in vitro fitness attenuation of the Cluster 5 virus in the Vero-E6 cell line. Potential alterations in antigenicity conferred by amino acid changes in the spike protein that include three substitutions (Y453F, I692V, and M1229I) and a loss of two amino acid residues 69 and 70 (ΔH69/V70), were evaluated in a virus microneutralization assay. Compared to a reference strain, the Cluster 5 variant showed reduced neutralization in a proportion of convalescent human COVID-19 samples. The findings underscore the need for active surveillance SARS-CoV-2 infection and virus evolution in susceptible animal hosts.

scholarly journals BAG-6 is essential for selective elimination of defective proteasomal substrates

2010 ◽  
Vol 190 (4) ◽  
pp. 637-650 ◽  
Author(s):  
Ryosuke Minami ◽  
Atsuko Hayakawa ◽  
Hiroki Kagawa ◽  
Yuko Yanagi ◽  
Hideyoshi Yokosawa ◽  
...  
Keyword(s):  
Mhc Class I ◽  
Mammalian Cells ◽  
Human Major Histocompatibility Complex ◽  
Histocompatibility Complex ◽  
Selective Elimination ◽  
Nascent Chain ◽  
Vitro Analysis ◽  
In Vitro Analysis

BAG-6/Scythe/BAT3 is a ubiquitin-like protein that was originally reported to be the product of a novel gene located within the human major histocompatibility complex, although the mechanisms of its function remain largely obscure. Here, we demonstrate the involvement of BAG-6 in the degradation of a CL1 model defective protein substrate in mammalian cells. We show that BAG-6 is essential for not only model substrate degradation but also the ubiquitin-mediated metabolism of newly synthesized defective polypeptides. Furthermore, our in vivo and in vitro analysis shows that BAG-6 interacts physically with puromycin-labeled nascent chain polypeptides and regulates their proteasome-mediated degradation. Finally, we show that knockdown of BAG-6 results in the suppressed presentation of MHC class I on the cell surface, a procedure known to be affected by the efficiency of metabolism of defective ribosomal products. Therefore, we propose that BAG-6 is necessary for ubiquitin-mediated degradation of newly synthesized defective polypeptides.

Hair cell development in vivo and in vitro: Analysis by using a monoclonal antibody specific to hair cells in the chick inner ear

2002 ◽  
Vol 445 (2) ◽  
pp. 176-198
Author(s):  
Kenji Kondo ◽  
Hiroshi Sagara ◽  
Kazushige Hirosawa ◽  
Kimitaka Kaga ◽  
Satsuki Matsushima ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Hair Cell ◽  
Inner Ear ◽  
Hair Cells ◽  
Cell Development ◽  
Vitro Analysis ◽  
In Vitro Analysis

scholarly journals Structural Modelling and in-silico characterization of a Novel Thermophilic β-amylase from Clostridium thermosulfuregenes

2021 ◽  
pp. 1-22
Author(s):  
Shafiqa Nayel ◽  
Mohd Shahir Shamsir ◽  
Farid Ahmad Danishfar
Keyword(s):  
Amino Acid ◽  
Binding Sites ◽  
Hydrolytic Enzyme ◽  
Beta Sheets ◽  
Amino Acid Residues ◽  
Structural Domains ◽  
Structural Conformation ◽  
In Silico Characterization

β-amylase is a hydrolytic enzyme that is involved in breaking down starch and producing energy. Since the discovery of β-amylase, it has been applied in various applications especially in the food industry. In this study, a novel β-amylase from Clostridium thermosuluregen, a thermophilic anaerobic bacterium that ferments its extracellular emulsion to ethanol at 62 ℃ was modelled and studied using bioinformatics tools and compared with B. cereus β-amylases that functions at mesophilic conditions. The results showed that the overall structural conformations, secondary structures, and important residues involved in active and binding sites were identified in both proteins. The results revealed that the modelled β-amylase of C. thermosulfuregen is very similar with respect to the global conformation, location of active and binding sites. Both proteins showed identical structural domains with the thermophilic variant possessing a high percentage of hydrophobic amino acid residues, polar amino acid residues, and differences in secondary composition such as loops and beta sheets as the potential evolutionary thermal adaptations that make it stable enzyme that functions up to 70 ℃. The results suggest that the thermal stability are not dependent on one single unique mechanism and may use one or a combination of the mechanisms to sustain its structural conformation at a higher operating temperature. Overall, considering the common properties of this modelled protein with the β-amylase of B. cereus, it can be assumed that if the β-amylase of C. thermosulfuregen were expressed in-vitro, it would produce a stable protein that possesses the hydrolysis function for C. thermosulfuregen to break down the starch and sugar formation.

scholarly journals The isolation and characterization of corticotropin from the pituitary gland of the ostrich Struthio camelus

1977 ◽  
Vol 165 (3) ◽  
pp. 519-523 ◽  
Author(s):  
R J Naudé ◽  
W Oelofsen
Keyword(s):  
Amino Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Amino Acid Residues ◽  
Struthio Camelus ◽  
Isolation And Characterization ◽  
Isoelectric Points ◽  
Pituitary Glands ◽  
Amide Content

1. Avian corticotropin (ACTH) was purified from both fresh and aged pituitary glands of the ostrich Struthio camelus. 2. The isolation of corticotropin in pure form involved acid/acetone extraction, NaCl fractionation, CM-cellulose chromatography and Sephadex G-50 chromatography. 3. The hormone preparations from fresh and aged glands behaved as single substances on polyacrylamide-gel electrophoresis, and both preparations were found to consist of 39 amino acid residues, in identical molar proportions for the different amino acids. 4. The isoelectric points of the two hormone preparations were estimated to be in the range pH 8.3-8.7, indicating possible differences in amide content, and the N-terminal amino acid of both preparations appeared to be serine. 5. The hormone preparations from fresh and aged glands exhibited similar biological potencies (73 and 77 i.u./mg respectively), as measured by steroidogenesis in vitro. 6. Apart from possible differences in amide content, the corticotropin preparations obtained from fresh and aged glands appear to be indistinguishable.

scholarly journals Skeletal Characterization of Smurf2-Deficient Mice and In Vitro Analysis of Smurf2-Deficient Chondrocytes

PLoS ONE ◽  
2016 ◽  
Vol 11 (1) ◽  
pp. e0148088 ◽  
Author(s):  
Henry Huang ◽  
Eric S. Veien ◽  
Hong Zhang ◽  
David C. Ayers ◽  
Jie Song
Keyword(s):  
Vitro Analysis ◽  
Deficient Mice ◽  
In Vitro Analysis
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