By Nathan P. Colowick, Nathan P. Kaplan, C. H.W. Hirs, Serge N. Timasheff
The significantly acclaimed laboratory regular, Methods in Enzymology, is without doubt one of the so much hugely revered courses within the box of biochemistry. considering that 1955, each one quantity has been eagerly awaited, often consulted, and praised via researchers and reviewers alike. The sequence includes a lot fabric nonetheless appropriate at the present time - actually a vital booklet for researchers in all fields of lifestyles sciences
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Extra info for Enzyme Structure Part E
0. 0. Digestion is carried out at 50 ° for 10 hr. 0, as before. To these combined supernatants the following bound enzymes are added: APM (600 mg, damp),Pronase (500 mg, damp), and prolidase ( 1 g, damp). The digestion is done at 37 ° for 12 hr. The digest is treated with additional buffer as described above. 5 ml is applied to an amino acid analyzer column. The remaining sample is hydrolyzed with 6 N HC1 (Pierce) in an evacuated sealed tube at 110 ° for 24 hr. Discussion. PROLINE RELEASE. CPY readily releases proline except when it is adiacent to glycine.
Although not yet widely employed, reagents other than ninhydrin (like f l u o r e s c a m i n e a n d o - p h t h a l a l d e h y d e ) h a v e been i n t r o d u c e d in t h e p a s t few y e a r s to d e t e c t a m i n o acids in i o n - e x c h a n g e column effluents. Use of the i n t e r n a l s t a n d a r d s d e s c r i b e d in this r e p o r t in a n a l y z e r s e q u i p p e d w i t h such d e t e c t i o n m e t h o d s h a s y e t to be r e p o r t e d . H e n c e t h e i r s u i t a b i l i t y for this p u r p o s e r e m a i n s to be e s t a b l i s h e d .
Mathews, P. G. H. Byfield, and I. MacIntyre, J. Chromatogr. I10, 369 (1975). C. L. Zimmerman, J. J. Pisano, and E. Appella, Biochem. Biophys. Res. Commun. 55, 1220 (1973). 46 AMINO ACID ANALYSIS  especially useful for the analysis of phenylthiohydantoins (PTH) and of 2,4-dinitrophenyl (DNP) and 1-dimethylaminonaphthalene-5-sulfonyl (dansyl) amino acids. Materials and M e t h o d Several manufacturers offer suitable instruments. TM A gradient maker is recommended because it shortens the analysis.
Enzyme Structure Part E by Nathan P. Colowick, Nathan P. Kaplan, C. H.W. Hirs, Serge N. Timasheff