Traut

Traut TW (1994) Physiological concentrations of purines and pyrimidines. Mol Cell Biochem 140 1-22... 

Williams, D.B.G., Traut, T, Kriel, F.H. and van Zyl, W.E. (2007) Bidentate amino-and iminophosphine ligands in mono-and dinuclear gold(I) complexes Synthesis, structures and AuCl displacement by AuC E . Inorganic Chemistry Communications, 10(5), 538-542. 

Traut, E. F. and E. W. Passarelli, Placebos in the Treatment of Rheumatoid Arthritis and Other Rheumatic Conditions , Annals of the Rheumatic Diseases 16 (1957) 18-22... 

Modification of Amines with 2-lminothiolane (Traut s Reagent)... 

At high pH (10.0), Traut s reagent also is reactive with aliphatic and aromatic hydroxyl groups, although the rate of reaction with these groups is only about 0.01 that of primary... 

Figure 1.61 Traut s reagent can undergo side reactions after the modification of an amine-containing molecule. The terminal thiol group can recyclize to create another iminothiolane derivative that effectively ties up the thiol.

Dissolve the Traut s reagent (Thermo Fisher) in water at a concentration of 2mg/ml (makes a 14.5 mM stock solution). The solution should be used immediately. For the modification of IgG at a concentration of lOmg/ml using a 10-fold molar excess of Traut s reagent, add 45.8 pi of the stock solution to each ml of the protein solution. 

Purify the thiolated protein from unreacted Traut s reagent by gel filtration using your buffer of choice (i.e., 20mM sodium phosphate, 0.15M NaCl, ImM EDTA, pH 7.2). The addition of EDTA to this buffer helps to prevent oxidation of the sulfhydryl groups and the resultant disulfide formation. After purification, use the thiolated protein immediately... 

The number of sulfhydryls created on the immunoglobulin using thiolation procedures such as this one is more critical to the yield of conjugated enzyme molecules than the molar excess of maleimide-activated enzyme used in the conjugation reaction. Therefore, it is important to use a sufficient excess of Traut s reagent to obtain a sufficient number of available sulfhydryls. 

Add 2-iminothiolane (Thermo Fisher) to this solution to give a molar excess of 20-40 X over the amount of antibody present (MW of Traut s reagent is 137.63). Addition of solid 2-iminothiolane may be done despite the fact that the compound is relatively insoluble in aqueous solution. As the reagent reacts, it will be completely drawn into solution. Alternatively, a stock solution of Traut s may be made in DMF and an aliquot added to the antibody solution (not to exceed 10 percent DMF in the final solution). 

Cover, J.A., Lambert, J.M., Norman, C.M., and Traut, R.R. (1981) Identification of proteins at the subunit interface of the Escherichia coli ribosome by cross-linking with dimethyl 3,3 -dithiobis(propionimi date). Biochemistry 20, 2843-2852. 

Sun, T.T., Bollen, A., Kahan, L., and Traut, R.R. (1974) Topography of ribosomal proteins of the Escherichia coli 30S subunit as studied with the reversible cross-linking reagent methyl 4-mercaptobu-tyrimidate. Biochemistry 13, 2334—2340. 

Traut, R.R., Casiano, C., and Zecherle, N. (1989) Cross-linking of protein subunits and ligands by the introduction of disulfide bonds. In Protein Function—A Practical Approach (T.E. Creighton, ed.), pp. 101-133. IRL Press at Oxford University, Oxford. 

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