Derivatization of Cysteine and Cystine for Fluorescence Amino Acid Analysis with the &Phthaldiald&hyde/Z-Mercaptoethanol Reagent*

نویسندگان

  • Kang S. Lee
  • Dennis G. Drescher
چکیده

Previous reports (Drescher, D. G., and Lee, K. S. (1978) Anal. Biochem. 84, 559-569; Lee, K. S., and Drescher, D. G. (1978) Int. J. Biochem. 9,457-467) have shown that high performance liquid chromatographic analysis of amino acids with the o-phthaldialdehyde/2mercaptoethanol reagent (OPA/P-ME) is one of the most sensitive procedures currently available for micro amino acid analysis. In the present paper, methods are presented for the modification of cysteine and cystine in proteins for micro amino acid analysis using OPA/2ME. Cysteine and cystine, which both show low fluorescence with OPA/P-ME, are converted to cysteic acid with performic acid directly, or to S-3-sulfopropylcysteine with 1,3-propane sultone after reduction of cystine with tri-n-butylphosphine. Cysteic acid and S-3sulfopropylcysteine form highly fluorescent adducts with OPA/S-ME. The formation of S-3-sulfopropylcysteine in proteins and the subsequent hydrolysis of the proteins with methanesulfonic acid are particularly useful for complete amino acid analysis at the picomole level using a single sample.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Derivatization of cysteine and cystine for fluorescence amino acid analysis with the o-phthaldialdehyde/2-mercaptoethanol reagent.

Previous reports (Drescher, D.G., and Lee, K.S. (1978) Anal. Biochem. 84, 559-569; Lee, K.S., and Drescher, D.G. (1978) Int. J. Biochem. 9, 457-467) have shown that high performance liquid chromatographic analysis of amino acids with the o-phthaldialdehyde/2-mercaptoethanol reagent (OPA/2-ME) is one of the most sensitive procedures currently available for micro amino acid analysis. In the prese...

متن کامل

A Modified Amino Acid Analysis Using PITC Derivatization for Soybeans with Accurate Determination of Cysteine and Half-Cystine

Breeding efforts to change the amino acid profile of seed protein and the assessment of genetic variation for amino acid composition among soybean germplasm resources have been hampered by lack of a rapid and inexpensive method for amino acid determination. A modified procedure presented here is partly based on a gasphase hydrolysis and precolumn derivatization for HPLC analysis. The procedure ...

متن کامل

The chromatographic determination of cystine and cysteine residues in proteins as s-beta-(4-pyridylethyl)cysteine.

Disullide bonds in proteins and in seed meals were reduced with /3-mercaptoethanol to sulfhydryl groups. The generated sulfhydryl groups were then alkylated by treatment with 4-vinylpyridine. The cysteine residues were thus derivatized to S-(4-pyridylethyl)-L-cysteine residues. The cysteine derivative is stable to acid hydrolysis and on amino acid analysis by ion exchange elutes just before arg...

متن کامل

Analysis of Plasma Amino Acids Using RP-HPLC and Pre-Column Derivatization with OPA/3-MPA

ABSTRACT        Background and Objective: Measurement of amino acids is an important tool for metabolic studies and evaluation of patients’ clinical condition. The aim of this study was to analyze the plasma amino acids using reverse phase high performance liquid chromatography techniques (RP-HPLC) with pre-column derivatization by o-phthalaldehyde (OPA)...

متن کامل

Mechanism of growth stimulation of L1210 cells by 2-mercaptoethanol in vitro. Role of the mixed disulfide of 2-mercaptoethanol and cysteine.

The mechanism of the growth-promoting action of 2-mercaptoethanol on mouse lymphoma L1210 cells in vitro has been studied. The normal line of L1210 is deficient in its capacity to take up cystine, and this limits the growth of the cells in normal culture media containing cystine. In the presence of 2-mercaptoethanol, the cells obtain a high capacity to utilize cystine in the medium. The reactio...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره   شماره 

صفحات  -

تاریخ انتشار 2002