By J. S. Davies
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Extra info for Amino Acids, Peptides and Proteins (SPR Amino Acids, Peptides, and Proteins (RSC)) (Vol 2)
I. Harris, B. S. Hartley, and R. Leberman, Biochem. , 1969,112,679. 112 L. King and R. N. Perham, unpublished work. 113 M. H. Freedman, A. L. Grossberg, and D. Pressman, Biochemistry, 1968, 7, 1941. 2 It has also been pointed o u t 2 that unblocked sulphydryl groups might add irreversibly across the double bond of both maleic and 2-methylmaleic (citraconic) anhydride. A study of the reaction of aldolase with citraconic anhydride 114has revealed that although the rate of alkylation of thiol groups is slow under the conditions used for acylation of protein amino-groups (possibly because of steric hindrance by the substituent methyl group at the double bond compared with maleic anhydride), irreversible loss of protein thiol groups does occur and is almost certainly the reason for the subsequent difficulties in regenerating enzyme activity.
At the other end of the scale, experiments have been described on the DNA-dependent RNA polymerase from E. 5 pg of ~ r 0 t e i n . l ~ ~ Such efforts merit close attention. The most useful single advance recently has probably been the introduction of the sodium dodecyl sulphate (SDS)-gel electrophoresis The presence of large amounts of SDS in the gels ensures that all polypeptide chains are strongly denatured and, presumably because the peptide chains are saturated with bound SDS, a linear relatioiiship is found to hold between the electrophoretic mobility and the log (molecular weight) of the peptide chain, analogous to the gel filtration system referred to above.
It is sometimes, though fortunately rarely, found that an Edman degradation refuses to go through an Asp-Gly or Asn-Gly sequence and it has been suggested 84 that the lack of a side-chain in glycine might be a contributing factor in allowing the Asx-Gly sequence to form the cyclic imide and thence the more stable p-aspartyl linkage, thereby inhibiting the degradation (although it must be confessed that this does not explain why the rearrangement happens only in isolated cases). It is 78 79 82 R3 84 P.