Cyanogen bromide cleavage of proteins in salt and buffer solutions.
Protocols for recombinant polypeptide production should provide high yields and be efficient, user friendly, and time saving. To perform cyanogen bromide (CNBr) cleavage of fusion proteins, the majority of researchers first desalted and vacuum-dried samples and then dissolved them in aqueous formic or trifluoroacetic acid. We propose to exclude the desalting step and run CNBr cleavage directly. We show that the commonly used Tris-HCl, sodium phosphate, NaCl, imidazole, and guanidine-HCl do not interfere with the reaction under acidic conditions. Omitting the desalting step does not decrease the final yields of target products, as demonstrated for fusion proteins of different origin and composition.
Andreev
Yaroslav A
YA
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya, 16/10, 117997 Moscow, Russia. ay@land.ru
Kozlov
Sergey A
SA
Vassilevski
Alexander A
AA
Grishin
Eugene V
EV
D002021:Buffers; D003488:Cyanogen Bromide / Q000737:chemistry*; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D012492:Salts / Q000737:chemistry*; D013879:Thioredoxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D002021:Buffers; D011993:Recombinant Fusion Proteins; D012492:Salts; D013879:Thioredoxins; D003488:Cyanogen Bromide