S. Cohen, Kathryn DeAntonis, D. Michaud
1993
Citations
2
Influential Citations
43
Citations
Journal
Techniques in Protein Chemistry
Abstract
Publisher Summary This chapter discusses compositional protein analysis using 6-aminoquinolyl- N -hydroxysuccinimidyl carbamate (AQC)—a novel derivatization reagent. Today many standard protein chemistry protocols—such as Edman degradation and peptide mapping—can be accomplished with low or even subpicomole sample amounts through the use of modem liquid chromatography. Amino acid analysis has been assisted in this drive for higher sensitivity by the use of an array of precolumn derivatization reagents that yield easily detected labels. The synthesis of a novel activated carbamate, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC), forms the basis for a precolumn derivatization method and provides very accurate amino acid analysis of hydrolyzed peptides and proteins. A rapid reaction with amino acids forms highly stable asymmetric urea compounds with good fluorescence characteristics. A dramatic shift in fluorescence emission maximum for the amino acid adducts compared to the major reagent peak allows the direct injection of the sample reaction mixture with no interference from excess reagent. The chapter describes the application of AQC to the analysis of protein and peptide hydrolyzate samples, with an emphasis on the accuracy and reproducibility of the compositional data.