TY - JOUR
T1 - Carbodiimide-mediated O-sulfation of hydroxy-amino acids and peptides
T2 - A reaction suitable for radiolabeling
AU - Pongor, Sandor
AU - Brownlee, Michael
AU - Cerami, Anthony
N1 - Funding Information:
We thank Dr. Stanford Moore and Dr. Peter Blackburn for valuable discussions. We also thank Dr. Peter Ulrich for suggesting TFMS as a protonating agent. The skilled technical assistance of Maria Pospischil and Seth P. Monkarsh is gratefully acknowledged. The expert secretarial assistance of Ms. Alice Striegel is greatly appreciated. This work was supported in part by NIH Grant R01 AM19655, and a generous gift from Mr. Howard Dahl, Lacrosse, Wisconsin. Dr. Brownlee is the recipient of Public Health Service Special Emphasis Research Career Award l-KOl-AM00589-01 SRC from the National Institutes of Arthritis, Metabolism, and Digestive Diseases, and the National Heart, Lung, and Blood Institute.
PY - 1985/5/1
Y1 - 1985/5/1
N2 - Carbodiimide-mediated sulfation of hydroxy-amino acids, peptides, and proteins can be accomplished in dry dimethylformamide by incubation in a 20-50 molar excess of sulfuric acid and various concentrations of dicyclohexyl carbodiimide [(1-ethyl-(3-dimethylaminopropyl)carbodiimide or 1-cyclohexyl-3-(2-morpholoethyl)carbodiimide p-toluene sulfonate)] at 4 °C for 2-4 h. Under these conditions, hydroxy-amino acids are quantitatively converted into O-sulfates, while cysteine yields the S-sulfonate. Other amino acids, including tryptophan, do not react and are recovered quantitatively. Porcine sodium insulin yields a product that can be separated into six bands by nondenaturing polyacrylamide gel electrophoresis. Radioloabeling of peptides by this method can be carried out with a high degree of efficiency if the added [35S]sulfuric acid is used carrier free with an acid excess provided by trifluoromethyl sulfonic acid. Under these conditions, over 60% of [35S]sulfuric acid was incorporated into insulin and bovine serum albumin. This method may prove useful in the radiolabeling of other peptides and proteins.
AB - Carbodiimide-mediated sulfation of hydroxy-amino acids, peptides, and proteins can be accomplished in dry dimethylformamide by incubation in a 20-50 molar excess of sulfuric acid and various concentrations of dicyclohexyl carbodiimide [(1-ethyl-(3-dimethylaminopropyl)carbodiimide or 1-cyclohexyl-3-(2-morpholoethyl)carbodiimide p-toluene sulfonate)] at 4 °C for 2-4 h. Under these conditions, hydroxy-amino acids are quantitatively converted into O-sulfates, while cysteine yields the S-sulfonate. Other amino acids, including tryptophan, do not react and are recovered quantitatively. Porcine sodium insulin yields a product that can be separated into six bands by nondenaturing polyacrylamide gel electrophoresis. Radioloabeling of peptides by this method can be carried out with a high degree of efficiency if the added [35S]sulfuric acid is used carrier free with an acid excess provided by trifluoromethyl sulfonic acid. Under these conditions, over 60% of [35S]sulfuric acid was incorporated into insulin and bovine serum albumin. This method may prove useful in the radiolabeling of other peptides and proteins.
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U2 - 10.1016/0003-9861(85)90187-0
DO - 10.1016/0003-9861(85)90187-0
M3 - Article
C2 - 3994383
AN - SCOPUS:0021853138
VL - 238
SP - 458
EP - 463
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
SN - 0003-9861
IS - 2
ER -