scholarly journals A new method for the selective isolation of cysteine-containing peptides. Specific labelling of the thiol group with a hydrophobic chromophore

1983 ◽  
Vol 211 (1) ◽  
pp. 163-171 ◽  
Author(s):  
J Y Chang ◽  
R Knecht ◽  
D G Braun
Keyword(s):  
High Pressure ◽  
Light Chain ◽  
Visible Region ◽  
High Sensitivity ◽  
Thiol Group ◽  
Reversed Phase ◽  
New Method ◽  
Kappa Light Chain ◽  
Selective Isolation ◽  
Specific Labelling

A new method for the selective isolation of cysteine-containing peptides was designed. The method is based on the specific labelling of thiol groups with a hydrophobic chromophore followed by enzymic fragmentation of the labelled protein and reversed-phase high-pressure liquid-chromatographic separation of the peptide mixture. This new method has several distinct advantages: (1) the hydrophobic-chromophore-labelled cysteine-containing peptides are easily separated from non-cysteine-containing peptides by reversed-phase high-pressure liquid chromatography; (2) only cysteine-containing peptides are detected in the visible region with sensitivity at the low picomole level; this high sensitivity allows isolation of nanogram amounts of pure cysteine-containing peptide; (3) during sequence determination of the chromophore-labelled cysteine-containing peptides, the cysteine residues are released as coloured anilinothiazolinone derivatives and can be detected directly in the picomole range; (4) with proteins bearing several disulphide groups, each disulphide group may undergo a different degree of reduction, and therefore the recovery of individual cysteine-containing peptides may be used to deduce the disulphide linkages present in the native protein. Two thiol-specific reagents, 4-dimethylaminoazobenzene-4′-iodoacetamide and 4-dimethylaminoazobenzene-4′-N-maleimide, were synthesized and characterized. The method was successfully used to isolate five cysteine-containing peptides from a completely reduced monoclonal-antibody kappa-light chain raised against the azobenzenearsonate determinant and six cysteine-containing peptides from a kappa-light chain raised against streptococcal group A polysaccharide. The principle of this method is applicable to the isolation of any peptide containing amino acid residues that can be specifically labelled with a hydrophobic chromophore.

scholarly journals N-terminal sequence analysis of polypeptide at the picomole level

1981 ◽  
Vol 199 (3) ◽  
pp. 557-564 ◽  
Author(s):  
J Y Chang
Keyword(s):  
High Pressure ◽  
Sequence Analysis ◽  
Visible Region ◽  
Reversed Phase ◽  
Complete Sequence ◽  
Complete Analysis ◽  
Phenyl Isothiocyanate ◽  
Terminal Sequence ◽  
Baseline Noise ◽  
Liquid Chromatographic

This paper describes a manual method for N-terminal sequence analysis of polypeptides at subnanomole sensitivity. The polypeptide is degraded stepwise by using the dimethylaminoazobenzene isothiocyanate/phenyl isothiocyanate double-coupling method, and the released dimethylaminoazobenzenethiohydantoins of amino acids were identified by reversed-phase high-pressure liquid chromatography. The dimethylaminoazobenzenethiohydantoins are coloured compounds and can be detected in the visible region with the sensitivity limit of 1 pmol (signal-to-baseline noise ratio 5). A high-pressure liquid-chromatographic method was developed for complete analysis of all amino acid dimethylaminoazobenzenethiohydantoin derivatives, including the by-products of serine and threonine. Thus, without use of an automatic sequenator or radioactive materials, it is possible to determine the complete sequence of peptides and N-terminal sequence of proteins with less than 1 nmol of material.

Inactivation of Nf-κb Pathway by Taxifolin Attenuates Sepsis-Induced Acute Lung Injury

2019 ◽  
Vol 18 (2) ◽  
pp. 176-182
Author(s):  
Chen Weiyan ◽  
Deng Wujian ◽  
Chen Songwei
Keyword(s):  
Acute Lung Injury ◽  
B Cells ◽  
Lung Injury ◽  
Signaling Pathway ◽  
Light Chain ◽  
Cecal Ligation ◽  
Nuclear Factor ◽  
Kappa Light Chain ◽  
Cecal Ligation And Puncture ◽  
Light Chain Enhancer

Acute lung injury is a clinical syndrome consisting of a wide range of acute hypoxemic respiratory failure disorders. Sepsis is a serious complication caused by an excessive immune response to pathogen-induced infections, which has become a major predisposing factor for acute lung injury. Taxifolin is a natural flavonoid that shows diverse therapeutic benefits in inflammation- and oxidative stress-related diseases. In this study, we investigated the role of taxifolin in a mouse model of cecal ligation and puncture-induced sepsis. Cecal ligation and puncture-operated mice presented damaged alveolar structures, thickened alveolar walls, edematous septa, and hemorrhage compared to sham-treated controls. Cecal ligation and puncture mice also showed increased wet-to-dry (W/D) lung weight ratio and elevated total protein concentration and lactate dehydrogenase level in bronchoalveolar lavage fluid. Taxifolin treatment protected animals against sepsis-induced pulmonary damage and edema. Septic mice presented compromised antioxidant capacity, whereas the administration of taxifolin prior to cecal ligation and puncture surgery decreased malondialdehyde concentration and enhanced the levels of reduced glutathione and superoxide dismutase in mice with sepsis-induced acute lung injury. Moreover, cecal ligation and puncture-operated mice showed markedly higher levels of proinflammatory cytokines relative to sham-operated group, while taxifolin treatment effectively mitigated sepsis-induced inflammation in mouse lungs. Further investigation revealed that taxifolin suppressed the activation of the nuclear factor kappa-light-chain-enhancer of activated B cells signaling pathway in cecal ligation and puncture-challenged mice by regulating the phosphorylation of p65 and IκBα. In conclusion, our study showed that taxifolin alleviated sepsis-induced acute lung injury via the inhibition of nuclear factor kappa-light-chain-enhancer of activated B cells signaling pathway, suggesting the therapeutic potential of taxifolin in the treatment sepsis-induced acute lung injury.

scholarly journals Untranslated immunoglobulin kappa light chain mRNA in a lambda light chain-producing mouse myeloma, MOPC104E.

1980 ◽  
Vol 255 (11) ◽  
pp. 5291-5295
Author(s):  
T. Kataoka ◽  
M. Ono ◽  
M. Kawakami ◽  
Y. Ikawa ◽  
M. Aida ◽  
...  
Keyword(s):  
Light Chain ◽  
Kappa Light Chain ◽  
Lambda Light Chain ◽  
Immunoglobulin Kappa ◽  
Mouse Myeloma

A Murine Monoclonal Multireactive Immunoglobulin Kappa Light Chain

1994 ◽  
Vol 39 (1) ◽  
pp. 107-110 ◽  
Author(s):  
W. MAHANA ◽  
F. JACQUEMART ◽  
M. ERMONVAL
Keyword(s):  
Light Chain ◽  
Kappa Light Chain ◽  
Immunoglobulin Kappa

Quantification of Tire-Tread Particles Using Extractable Organic Zinc as Tracer

1999 ◽  
Vol 72 (5) ◽  
pp. 969-977 ◽  
Author(s):  
Patrik Fauser ◽  
Jens Christian Tjell ◽  
Hans Mosbaek ◽  
Kim Pilegaard
Keyword(s):  
Atomic Absorption Spectrometry ◽  
Atomic Absorption ◽  
Environmental Samples ◽  
High Sensitivity ◽  
Absorption Spectrometry ◽  
Soil Samples ◽  
New Method ◽  
Organic Zinc

Abstract A method for identifying and quantifying tire-tread particles in the environment has been developed. It is based on the measurement of extractable organic zinc. The high sensitivity of atomic absorption spectrometry (AAS) with a heated graphite atomizer (HGA) permits assessment of submilligram amounts of tire debris in environmental samples. The analysis is performed on aerosol and soil samples. This new method is more accurate and faster than the previously reported IR method.

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