Identification of phenylthiohydantoins of amino acids by thin-layer chromatography on a plastic-backed silica-gel plate

The complete amino acid sequence of ribosomal protein L34 has been established by improved micro techniques with 3 mg of the lyophilized protein. The protein was digested with trypsin, thermolysin and chymotrypsin and the resulting peptides were isolated from fingerprints performed on cellulose thin-layer plates. The amino acid sequences of the peptides were determined by the combined micro dansyl-Edman technique using 5 - 10 nmol per sample. Aspartic acid and glutamic acid were distinguished from their amides by use of the color reaction of ninhydrin with the respective amino acid phenylthiohydantoins.

The sequence determination of a protein in a micro scale: the sequence analysis of ribosomal protein L34 of Escherichia coli.

The major polypeptide chain of bovine β-crystallin, βBp, was fragmented by means of cyanogen bromide treatment and by enzymatic digestions. Manual and automated Edman degradation of the resulting peptides provided the complete amino acid sequence of the 0Bp chain. The N-terminal alanine residue was shown to be N-α-acetylated by mass spectrometry. The chain has a length of 204 residues and a calculated molecular weight of 23210. There is a considerable degree of homology between the N-terminal and C-terminal halves of the chain, presumably reflecting a tandem duplication of a shorter ancestral gene. The sequence of βBp is sufficiently related to that of γ-crystallin II to place these proteins in the same superfamily. No sequence relationship was found with the α-crystallin chains.

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1432-1033.1981.tb06434.x

Primary structure of the bovine beta-crystallin Bp chain. Internal duplication and homology with gamma-crystallin.

Bitis gabonica venom. The amino acid sequence of phospholipase A.

Publisher Summary This chapter discusses thin-layer chromatography (TLC) for the analysis of amino acids and derivatives. The qualitative analyses that can be done by TLC includes comparison of peptide and protein hydrolyzates, control of the course of hydrolysis, end-group determination, identification in sequence analysis, and also screening for inborn errors of amino acid metabolism. The chapter discusses the special requirements for TLC and the importance of careful standardization of the technique, including the following: quality of layer material, layer thickness and homogeneity, layer activation and relative humidity, amount of sample applied, narrow starting point or line, effective drying of the applied sample, quality of solvent (aging), chamber saturation, chamber volume and geometry, temperature, distances of starting point and solvent front from immersion line, and drying of the layer prior to TLC in the second dimension. A thin-layer consisting of two different materials, a narrow zone of a material A along a border of the plate and a material B on the rest of the plate area is useful for trapping unwelcome or disturbing substances at the starting point or to enable two dimensional TLC on different stationary phases.

Gilbert Roseau

Papers

Révélation colorée des spots de phénylthiohydantoi̋ne d'acides aminés