Histidine separation from arginine

On the basis of data presently available, one would conclude that phosphorylation of serine in proteins probably occurs when the serine residues are adjacent to or in close proximity (sequence-wise) to basic or acidic amino acid residues. In the tertiary structure it would be expected that these residues would be near the surface of the protein. Recently, it has been shown for a wide variety of O-phosphoryl proteins that protein substrates of intracellular protein kinases all have a common feature when the caseins are excluded in general, all phosphorylated sites are separated from either lysine or arginine (in two cases, histidine) by no more than two amino acid residues (103a). 

Flgvire 4.5 The influence of endcapping on peak shape and retention of soee PTH-anino acids using a reversed-phase separation system. Peak identification 1 PTH-histidine, 2 PTH-arginine and 3 PTH-valine. (Reproduced with permission from ref. 71. Copyright Preston Publications, Inc.)... 

It is obvious from the infrared studies to date that the effect of a given additive may vary depending on the protein (e.g., [8]), the presence of other additives and other specific solution conditions, e.g., pH [12]. Therefore, the structure of each dried protein in each formulation should be studied with FTIR spectroscopy. Unfortunately, this will not be possible with certain formulations. If albumin is used, then, as is the case with any physical measurement, it will not be possible to separate the albumin contribution to the data from that of the protein drug. If other compounds (e.g., PVP, arginine, histidine, glycine) that absorb strongly in the... 

Proton resonance spectra of denatured proteins consist of sharp peaks which correspond to a summation of resonances from individual residues [129]. In C spectra of denatured proteins, it is possible to distinguish all the carbon resonances of the aromatic side chains of histidine, phenylalanine, tyrosine and tryptophan, and separate resonances from alanine, arginine, glycine, isoleucine, leucine, threonine, valine and occasionally methionine [130]. (A natural abundance C spectrum of a 13 mM solution of lysozyme takes only 4 hr accumulation time using 20 mm sample tubes [131]). 

Fig. 4.4.19. Separation of amino acids on a cation-exchange resin and with post-column derivation with ninhydrin. B was obtained 50 analyses after A. Peak identification D, aspartic acid T. threonine S, serine E. glutamic acid P. proiine G, glycine A. alanine C. cysteine V, valine M. methionine I, isoleucine L. leucine NL. norleucine F. phenylalanine O. ornithine K, lysine NH,. ammonia H. histidine R. arginine W. tryptophan. Reprinted from Ref. 96 with permission.

Fig. 11.2.12. Normal phase separation of amino acids. Chromatographic conditions column, Zorbax NH2 (250 x 4.6 mm I.D.) mobile phase, 10 mM potassium phosphate, pH 4.3 (A), acetonitrile-water 50 7 (v/v) (B) flow rate, 2 ml/min temperature, 35 °C. Peaks 1, phenylalanine 2, leucine 3, isoleucine 4, methionine 5, tyrosine 6, valine 7, proline 8, alanine 9, hypro 10, threonine 11, glycine 12, serine 13, histidine 14, cysteine 15, arginine 16, lysine 17, hydroxylysine 18, glutamic acid 19, aspartic acid. Reproduced from Smolensk et al. (1983), with permission.

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