Fluorescent Detection of a Partially Unfolded Conformation of Beta-Lactoglobulin Using Squaraine Dyes

2014 ◽  
Vol 335 (1) ◽  
pp. 43-50
Author(s):  
V.B. Kovalska ◽  
M.Yu. Losytskyy ◽  
L.V. Reis ◽  
P.F. Santos ◽  
S.M. Yarmoluk
Keyword(s):  
Fluorescent Detection ◽  
Squaraine Dyes ◽  
Partially Unfolded ◽  
Beta Lactoglobulin

Conformation of eukaryotic ribosomal RNAs

1983 ◽  
Vol 41 ◽  
pp. 592-593
Author(s):  
M. Boublik ◽  
G. Thornton ◽  
G. Oostergetel ◽  
J.F. Hainfeld ◽  
J.S. Wall
Keyword(s):  
Molecular Weight ◽  
Mass Measurement ◽  
Carbon Film ◽  
Structural Complexity ◽  
Scanning Transmission Electron Microscope ◽  
Electron Microscopic ◽  
Pure Nitrogen ◽  
Freeze Dried ◽  
Scanning Transmission ◽  
Partially Unfolded

Understanding the structural complexity of ribosomes and their role in protein synthesis requires knowledge of the conformation of their components - rRNAs and proteins. Application of dedicated scanning transmission electron microscope (STEM), electrical discharge of the support carbon film in an atmosphere of pure nitrogen, and determination of the molecular weight of individual rRNAs enabled us to obtain high resolution electron microscopic images of unstained freeze-dried rRNA molecules from BHK cells in a form suitable for evaluation of their 3-D structure. Preliminary values for the molecular weight of 28S RNA from the large and 18S RNA from the small ribosomal subunits as obtained by mass measurement were 1.84 x 106 and 0.97 x 106, respectively. Conformation of rRNAs consists, in general, of alternating segments of intramolecular hairpin stems and single stranded loops in a proportion which depends on their ionic environment, the Mg++ concentration in particular. Molecules of 28S RNA (Fig. 1) and 18S RNA (not shown) obtained by freeze-drying from a solution of 60 mM NH+4 acetate and 2 mM Mg++ acetate, pH 7, appear as partially unfolded coils with compact cores suggesting a high degree of ordered secondary structure.

DIFFERENT COW BREEDS MILKABILITY DEPENDING ON KAPPA-CASEIN AND BETA-GLOBULIN GENOTYPES

2019 ◽  
Keyword(s):  
Milk Fat ◽  
Animal Husbandry ◽  
Holstein Cows ◽  
Maintenance System ◽  
Plant Feeding ◽  
Beta Globulin ◽  
Ayrshire Breed ◽  
The Common ◽  
Milking Machine ◽  
Beta Lactoglobulin

Results of the Ayrshire breed cows, Holstein cows and Holstein-Yaroslavl cross breed cows milk productiv-ity analysis in the same environment conditions and depending on the kappa-casein and beta-lactoglobulin genotypes are presented in this article. The animals are kept in the common environmental conditions of the LLC Farming firm “Pakhma», the Ayrshire breeding plant. Feeding of cows here is carried out according to detailed norms, concentrated feed averages 43%. The maintenance system is year-round stalling, milking is carried out in the milk line, the DeLaval milking machine is used, and labor-intensive processes in animal husbandry are mechanized. The farm has a milk processing shop. It was established that all the inspected cows (n=91) had a prevailing AA kappa-casein genotype – 75,8% on average. BB Ayrshire breed and Hol-stein-Yaroslavl cross breed genotypes were not established. AB and BB beta-lactoglobulin genotypes are seen on average with a frequency of 44%. AA beta-lactoglobulin genotype in Holstein cows breed was not found. The indicators of statistically reliable difference in milkability among all groups and genotypes was not estab-lished. A higher consistence of protein in the milk of B-allele kappa-casein type cows is evident in all groups with no difference between breed belonging. The complex variant of CSN3/LGB genotypes showed that ac-cording to ultimate milk fat and protein yield the most productive cows were:АВ/ВВ, АВ/АВ genotype Ayr-shire cows, АВ/АВ Holstein genotype cows, AB/BB genotype Holstein-Yaroslavl cross breed cows. Hence, the most efficient cows are those with B-allele variants.

REALIZATION OF PARENTAL INDICES OF COW PRODUCTIVITY IN DIFFERENT MILKING BREEDS AT BREEDING FARM "AGROFIRMA "PAKHMA"

2020 ◽  
Keyword(s):  
Milk Yield ◽  
Cattle Breed ◽  
Holstein Cattle ◽  
Milk Productivity ◽  
Cattle Breeds ◽  
Significant Difference ◽  
Production Indices ◽  
High Level ◽  
Beta Lactoglobulin ◽  
Percentage Protein

Combination of genotypes of main types’ genetic markers causes milk productivity in cattle. The article studies correlation between kappa casein genotypes (C3N3), beta-lactoglobulin (LGB), their complexes and milk production indices in cows bred under equal conditions at a breeding farm. The calculation of parental milk yield and fat indices between the three breeds showed a significant difference (P>0,999) of PCI of Ayrshire and Holstein cattle breed milk yield - 3395 kg, PCI of fat - 0,22%; between Holstein and Yaroslavl cattle breeds - 3200 kg, PCI offat - 0,16%. A significant difference of realization indices of protein between all three breeds was not stated. The highest realization ofparental milk yield index was stated - between Ayrshire and Yaroslavl cattle breeds- 113%, Holstein cattle breed - 88%. As for the fat and protein indices, all three cattle breeds showed a high realization of PCI from 98% to 109%. However, Holstein cattle breed showed the highest percentage: protein - 101%, fat - 109%. A significant difference of PCI realization of milk productivity in CSN3 and LGB genotypes was not stated. It was also stated that there is a tendency of high level parental indices realization when there is a higher number of B-allele variants in genotypes of cows.

Improved Stability of a Protein Vaccine Through Elimination of a Partially Unfolded State

2004 ◽  
Author(s):  
Colleen A. McHugh ◽  
Ralph F. Tammariello ◽  
Charles B. Millard ◽  
John H. Carra
Keyword(s):  
Protein Vaccine ◽  
Unfolded State ◽  
Improved Stability ◽  
Partially Unfolded

scholarly journals An LED-Driven AuNPs-PDMS Microfluidic Chip and Integrated Device for the Detection of Digital Loop-Mediated Isothermal DNA Amplification

Micromachines ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 177 ◽  
Author(s):  
Zengming Zhang ◽  
Shuhao Zhao ◽  
Fei Hu ◽  
Guangpu Yang ◽  
Juan Li ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Microfluidic Chip ◽  
Cross Flow ◽  
Flow Channel ◽  
Isothermal Amplification ◽  
Fluorescent Detection ◽  
Lamp Method ◽  
Biological Studies ◽  
Micro Cavity ◽  
Integrated Device

The sensitive quantification of low-abundance nucleic acids holds importance for a range of clinical applications and biological studies. In this study, we describe a facile microfluidic chip for absolute DNA quantifications based on the digital loop-mediated isothermal amplification (digital LAMP) method. This microfluidic chip integrates a cross-flow channel for droplet generation with a micro-cavity for droplet tiling. DNA templates in the LAMP reagent were divided into ~20,000 water-in-oil droplets at the cross-flow channel. The droplets were then tiled in the micro-cavity for isothermal amplification and fluorescent detection. Different from the existing polydimethylsiloxane (PDMS) microfluidic chips, this study incorporates gold nanoparticles (AuNPs) into PDMS substrate through silica coating and dodecanol modification. The digital LAMP chip prepared by AuNPs-PDMS combines the benefits of the microstructure manufacturing performance of PDMS with the light-to-heat conversion advantages of AuNPs. Upon illumination with a near infrared (NIR) LED, the droplets were stably and efficiently heated by the AuNPs in PDMS. We further introduce an integrated device with a NIR heating unit and a fluorescent detection unit. The system could detect HBV (hepatitis B virus)-DNA at a concentration of 1 × 101 to 1 × 104 copies/μL. The LED-driven digital LAMP chip and the integrated device; therefore, demonstrate high accuracy and excellent performance for the absolute quantification of low-abundance nucleic acids, showing the advantages of integration, miniaturization, cost, and power consumption.

scholarly journals Mechanism of the small ATP-independent chaperone Spy is substrate specific

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Rishav Mitra ◽  
Varun V. Gadkari ◽  
Ben A. Meinen ◽  
Carlo P. M. van Mierlo ◽  
Brandon T. Ruotolo ◽  
...  
Keyword(s):  
Relative Affinity ◽  
Complex Protein ◽  
Partially Unfolded ◽  
Substrate Proteins ◽  
Partially Unfolded States

AbstractATP-independent chaperones are usually considered to be holdases that rapidly bind to non-native states of substrate proteins and prevent their aggregation. These chaperones are thought to release their substrate proteins prior to their folding. Spy is an ATP-independent chaperone that acts as an aggregation inhibiting holdase but does so by allowing its substrate proteins to fold while they remain continuously chaperone bound, thus acting as a foldase as well. The attributes that allow such dual chaperoning behavior are unclear. Here, we used the topologically complex protein apoflavodoxin to show that the outcome of Spy’s action is substrate specific and depends on its relative affinity for different folding states. Tighter binding of Spy to partially unfolded states of apoflavodoxin limits the possibility of folding while bound, converting Spy to a holdase chaperone. Our results highlight the central role of the substrate in determining the mechanism of chaperone action.

scholarly journals Safety evaluation of whey derived beta-lactoglobulin, Lacprodan® BLG

2021 ◽  
Author(s):  
Marianne Dybdahl ◽  
David Benjamin Selesko ◽  
Ulla Ramer Mikkelsen
Keyword(s):  
Safety Evaluation ◽  
Beta Lactoglobulin
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