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Biomedical subjects

C Redfield

Publications and source records attributed to C Redfield.

41 records · Page 3Linked to original sources

Identification of glycine spin systems in 1H NMR spectra of proteins using multiple quantum coherences.

Double-quantum filtered COSY and triple-quantum filtered COSY techniques have been compared for the tripeptide Gly-Tyr-Gly and for human lysozyme. The insertion of a triple-quantum filter in the COSY experiment leads to dramatic spectral simplification in the fingerprint region of the spectrum and permits the specific identification of glycine spin systems in the complex 1H NMR spectra of proteins. The assignment of these peaks to glycine H alpha can be confirmed using 2D double-quantum correlated spectroscopy.

Glycine↗

Surface accessibility of aromatic residues in human lysozyme using photochemically induced dynamic nuclear polarization NMR spectroscopy.

Resonances in the photo-CIDNP spectrum of human lysozyme have been assigned to specific spin systems despite extensive spectral overlap using the two-dimensional photo-CIDNP COSY experiment. Five of the 12 tyrosine, tryptophan and histidine residues of human lysozyme are found to be accessible to flavin dye in solution. This result is in good agreement with surface accessibility calculations carried out on the human lysozyme crystal structure. When amino acid differences are considered the photo-CIDNP results obtained for human lysozyme are in good agreement with results obtained previously for hen lysozyme.

Amino Acids↗

Lysozyme: a major secretory product of a human colon carcinoma cell line.

One of the major proteins secreted by an established human colon adenocarcinoma cell line has been isolated in 25% yield from the serum-free medium in which the cells were grown and identified as lysozyme. Its purification was achieved by sequential steps of acidification, cation-exchange chromatography, and reversed-phase high-performance liquid chromatography. It was recognized to be a human lysozyme on the basis of its molecular weight (14 000), isoelectric point (10.5), amino acid composition, and enzymatic activity. Its identity with previously characterized human lysozymes was established by amino-terminal sequence, peptide composition, immunological properties, NMR, and crystallography. A 4-day, 7-L collection of conditioned medium contained 20.3 mg of secreted protein of which 4.9 mg or approximately 24% of the total was tumor-derived lysozyme. The intracellular level of lysozyme was approximately 18 ng per 10(6) carcinoma cells. The possible significance of these findings in regard to the malignant process and tumor maintenance is discussed.

Adenocarcinoma↗

Complete assignment of the 1H NMR spectrum of the aromatic residues of lysozyme.

Assignment of the nuclear magnetic resonance (NMR) spectrum of the 59 non-exchangeable protons of the 13 aromatic amino acid residues of lysozyme has been completed using a combination of selective spin decoupling and nuclear Overhauser experiments. The experimental chemical shift data were used to simulate the aromatic region of the NMR spectrum at 300 MHz and at 470 MHz. Excellent agreement with the experimental spectra was obtained and the simulations permitted more accurate chemical shift values and resonance linewidths to be obtained. The current set of assignments is compared with those of the assignments made previously. Evidence for the motional behaviour of the aromatic amino acid side chains is presented.

Animals↗