Re: solubility of dinitrophenyl-peptides
Tim Kingan (tkingan@citrus.ucr.edu)
Tue, 17 Feb 1998 17:22:18 -0800
>Date: Tue, 17 Feb 1998 17:14:03 -0800
>To:xiaohui chen <xhchen@biocserver.BIOC.CWRU.Edu>
>From:tkingan@citrus.ucr.edu (Tim Kingan)
>Subject:Re: solubility of dinitrophenyl-peptides
>
>It seems that Fred Sanger had a similar "problem" when using DNFB for
>end-group analysis of insulin (Sanger, Biochemical J. 39, 507, 1945).
>However, he was able to hydrolyze the insoluble material and extract the
>DNP-amino acids for analysis. I have used a similar reagent,
>difluorodinitrobenzene, for cross-linking peptides to proteins in a
>two-step procedure in which the peptide is first reacted with excess DFDNB
>in 0.1 M K phos/7 M guanidine HCl, pH 7.2 (HS Tager, Analyt. Biochem. 71,
>367 1976). I've never had a solubility problem, but perhaps your plans
>can't accommodate the GuHCl.
>
>Tim Kingan
>Dept. of Entomology
>U of California
>Riverside, CA 92521
>(909)787-4369
>
>>Dear ABRFers,
>>
>>I derivatized peptides (from trypsin and chymotrypsin digestion of protein,
>>i.e., hemoglobin) with dinitrofluorobenzene in water/dimethyl formamide
>>mixture. Most formed dinitrophenyl (DNP)-peptides precipitated. After
>>thiolysis to remove DNP-tag from Cys, His and Tyr, only DNP-NH- left.
>>However, there were still significant amount of pellets in the solution.
>>Does anyone know a better solvent to dissolve DNP-peptides? Thanks.
>>
>>Xiaohui Chen, Ph.D.
>>Department of Biochemistry
>>School of Medicine
>>Case Western Reserve University
>>2109 Adelbert Road
>>Cleveland, OH 44106
>>Phone: 216-368-3250
>>Fax: 216-368-3419
>>Email: xhchen@biocserver.cwru.edu
>