scholarly journals Development of indole chemistry to label tryptophan residues in protein for determination of tryptophan surface accessibility

2007 ◽  
Vol 16 (6) ◽  
pp. 1204-1213 ◽  
Author(s):  
Carol L. Ladner ◽  
Raymond J. Turner ◽  
Robert A. Edwards
Keyword(s):  
Surface Accessibility ◽  
Tryptophan Residues ◽  
Indole Chemistry

Studies of Glutamate Dehydrogenase: Chemical Modification and Quantitative Determination of Tryptophan Residues

1974 ◽  
Vol 43 (2) ◽  
pp. 319-325 ◽  
Author(s):  
Veit Witzemann ◽  
Rudolf Koberstein ◽  
Horst Sund ◽  
Ihab Rasched ◽  
Hans Jornvall ◽  
...  
Keyword(s):  
Chemical Modification ◽  
Quantitative Determination ◽  
Glutamate Dehydrogenase ◽  
Tryptophan Residues

Rapid Determination of Tryptophan in Intact Proteins by Derivative Spectrophotometry

1993 ◽  
Vol 76 (6) ◽  
pp. 1168-1173 ◽  
Author(s):  
Dimitrios J Fletouris ◽  
Nickos A Botsoglou ◽  
Georgios E Papageorgiou ◽  
Antonios J Mantis
Keyword(s):  
Rapid Determination ◽  
Suspended Material ◽  
Feed Ingredient ◽  
Intact Proteins ◽  
Sample Extract ◽  
Tryptophan Residues ◽  
Food And Feed ◽  
Fourth Derivative ◽  
Uv Absorption Spectrum

Abstract A method for rapid determination of total tryptophan in intact proteins was developed. Sample is homogenized with 0.1 M sodium hydroxide, and the homogenate, if it contains suspended material, is centrifuged. Tryptophan is directly quantified in sample extract on the basis of its fourth-derivative UV absorption spectrum. Protein hydrolysis and/or extract purification is not required because the fourth-derivative transformation of the conventional analytical band around 285.5 nm virtually eliminates interferences arising from both tyrosine and other UV-absorbing components. When pure proteins were analyzed by the method, the values of tryptophan residues found coincided well with literature data based on sequence analysis. The applicability of the method was also tested on several food and feed ingredient samples selected to include a range of protein content and a variety of protein sources. Owing to its simplicity and reliability, the method is particularly recommended for everyday analysis of a large number of samples.

Determination of the excited-state lifetimes of the tryptophan residues in barnase, via multifrequency phase fluorometry of tryptophan mutants

Biochemistry ◽  
1992 ◽  
Vol 31 (3) ◽  
pp. 711-716 ◽  
Author(s):  
K. Willaert ◽  
R. Loewenthal ◽  
J. Sancho ◽  
M. Froeyen ◽  
A. Fersht ◽  
...  
Keyword(s):  
Excited State ◽  
Phase Fluorometry ◽  
Tryptophan Residues ◽  
Excited State Lifetimes

Direct Photometric Determination of Globulin in Serum

1971 ◽  
Vol 17 (5) ◽  
pp. 358-362 ◽  
Author(s):  
Harry Goldenberg ◽  
Patricia A Drewes
Keyword(s):  
Glyoxylic Acid ◽  
Photometric Determination ◽  
Confidence Limits ◽  
Serum Globulin ◽  
Clin Pathol ◽  
Free Tryptophan ◽  
Tryptophan Residues ◽  
Fractionation Method ◽  
Salt Fractionation

Abstract A convenient method for direct photometric determination of total globulin in serum has been developed using a one-tube, one-reagent system. The analysis is based on a variation of the Hopkins-Cole reaction, in which glyoxylic acid condenses with the tryptophan residues present in globulin to produce a purple color. In the presence of copper and under the conditions of the analysis, color intensity is proportional to serum globulin concentration. The method is standardized with either a serum control or a suitable tryptophan derivative (e.g., acetyl-DL-tryptophan). There is little or no interference from free tryptophan, albumin, bilirubin, lipemia, or mild hemolysis. The precision of the method is ±3% (95% confidence limits), and results agree with those obtained by the salt fractionation method of Wolfson et al. [Amer. J. Clin. Pathol. 18, 723 (1948)]: t = -0.88; critical t = 2.02; 95% confidence limits.

A new double-labelling procedure for determination of amino acid composition: Application to bacteriorhodopsin

1978 ◽  
Vol 56 (6) ◽  
pp. 517-520 ◽  
Author(s):  
H. Kaplan ◽  
D. C. H. Cheng ◽  
G. Oda ◽  
M. Kates
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Mixture ◽  
Acid Mixture ◽  
Amino Acid Derivatives ◽  
Double Labelling ◽  
Tryptophan Residues ◽  
Labelling Procedure ◽  
Derivatives Of

A new double-labelling procedure for amino acid analysis which requires only routine chromatographic equipment is described. When 1-fluoro-2,4-dinitro[3H]benzene is reacted with a mixture of 14C-labelled amino acids followed by reaction with the same 14C-labelled amino acid mixture diluted with an unlabelled sample of amino acids, the 3H: 14C ratio in the resulting 2,4-dinitrophenyl (DNP) amino acid derivatives of the diluted sample will be increased in proportion to the quantity of unlabelled amino acid in the diluted sample. This procedure gave reliable results when applied to the known proteins insulin and lysozyme. The procedure is most advantageous when applied to amino acids which are unstable during acid hydrolysis or present in low molar fractions.When applied to the analysis of the bacteriorhodopsin in Halobacterium cutirubrum, this procedure showed the presence of one histidine residue and four tryptophan residues per mole protein but no cystine or cysteine; in general, the analyses obtained were consistent with those originally reported by Oesterhelt, D. and Stoeckenius, W. (1971) (Nature (London) New Biol. 233, 149–152) for bacteriorhodopsin of H. halobium.

Quantitative gas chromatography/mass spectrometry determination of C-mannosylation of tryptophan residues in glycoproteins

2004 ◽  
Vol 329 (2) ◽  
pp. 199-206 ◽  
Author(s):  
Jean-Pierre Zanetta ◽  
Alexandre Pons ◽  
Colette Richet ◽  
Guillemette Huet ◽  
Philippe Timmerman ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Gas Chromatography Mass Spectrometry ◽  
Chromatography Mass Spectrometry ◽  
Tryptophan Residues

Simultaneous determination of tyrosine and tryptophan residues in proteins by second-derivative spectroscopy

1982 ◽  
Vol 126 (2) ◽  
pp. 251-257 ◽  
Author(s):  
Luigi Servillo ◽  
Giovanni Colonna ◽  
Ciro Balestrieri ◽  
Raffaele Ragone ◽  
Gaetano Irace
Keyword(s):  
Simultaneous Determination ◽  
Second Derivative ◽  
Derivative Spectroscopy ◽  
Tryptophan Residues ◽  
Second Derivative Spectroscopy

scholarly journals Determination of the number and relative position of tryptophan residues in various albumins

1976 ◽  
Vol 159 (3) ◽  
pp. 529-533 ◽  
Author(s):  
R C Feldhoff ◽  
T Peters
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Colorimetric Method ◽  
Peptide Chain ◽  
Tryptophan Residue ◽  
Serum Albumins ◽  
Molecular Weights ◽  
Tryptophan Residues ◽  
Relative Migration

A technique is described by which both the numbers of tryptophan residues and their approximate locations in the peptide chain of a protein can be determined by cleavage with N-bromosuccinimide followed by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. The number of new peptide bands appearing in the gel is a function of the number of tryptophan residues, and the relative migration of the bands permits calculation of peptide molecular weights and an estimation of the positions of the tryptophan residues in the peptide chain. The technique uses a sample of about 0.5 mg and is suitable for any protein that contains a small number of tryptophan residues. These are the very specimens that are difficult to assay accurately for tryptophan by spectrophotometric or colorimetric methods. Tryptophan residues which are within about 20 residues of the ends of the peptide chain or of each other would not be detected. The specificity of the cleavage with N-bromosuccinimide was ascertained by utilizing human serum albumin, which is known to have a single tryptophan residue at position 214. The technique was then applied to a comparative study of the numbers and locations of tryptophans in the serum albumins of 16 species, namely 11 mammals, three birds and two amphibians. The number of tryptophan residues were confirmed by an independent colorimetric method. All of the mammalian albumins contained a tryptophan residue near position 213. The three avian albumins examined have no tryptophan. Frog and toad albumins contained two tryptophan residues, which appear to be situated at different positions from those in mammalian albumins.

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