By Milton T.W. Hearn

High-Performance Liquid Chromatography of Proteins and Peptides.

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Activated pancreatic juice). However, these procedures are time consuming. The present paper describes a simple and rapid procedure allowing purification by reverse-phase HPLC of commercial trypsin and chymotrypsin in a small amount which is sufficient for protein-sequencing studies. Trypsin and chymotrypsin have been essential tools in the analysis of protein sequences. It is, however, always difficult to obtain from commercial sources entirely pure enzyme, either trypsin or chymotrypsin, which lacks contamination with the other.

1) from human liver, once thought to be a single molecular species, is actually composed of numerous molecular forms. More than 20 different isoenzymes (1-5) that exhibit a wide range of mobilities on starch-gel electrophoresis, even though they share many structural features, are now known. Thus, all ADH isoenzymes are dimeric, each monomer having a molecular weight of about 40,000, with a blocked TV-terminal amino acid, containing two zinc atoms and interacting with 1 mol of NAD(H) (6). However, while they also share many functional properties, there are distinctive differences.

Biophys. Acta 577, 159-169. 13. Jeffereys, A. , and Flavell, R. A. (1977) Cell 12, 429-439. 14. Hanash, S. , and Bunnell, K. (1981) in Advances in Hemoglobin Analysis (Hanash, S. , and Brewer, G. ), pp. 53-67, Alan R. Liss, New York. 15. Huisman, T. H. , Gardiner, M. , and Wilson, J. B. (1981) in Advances in Hemoglobin Analysis (Hanash, S. , and Brewer, G. ), pp. 6982, Alan R. Liss, New York. 16. Dozy, A. , Kan, Y. , Embury, S. , Mentzer, W. C , Yang, W. , Davis, J. , and Koenig, H. M. (1979) Nature (London) 280, 605-607.

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