Evidence for interaction of substrate analogs with chicken eggwhite lysozyme after exhaustive reduction of disulfide bonds.
In a study of factors that influence the remaining secondary structure of reduced chicken eggwhite lysozyme, N-acetyl-D-glucosamine (NAG) and N,N'-diacetylchitobiose (di-NAG) were found to alter the circular dichroic (CD) spectrum of the reduced protein and its carboxymethyl derivative (Cml). Thus, negative ellipticities in the far u.v. were greater in the presence of the analogs, with NAG being the more effective. For Cml, curve fitting analysis of the CD data indicated an increased helical content in the presence of NAG by an average of 3% of the chain length, while beta-structure decreased by an equivalent amount. Other compounds structurally related to NAG produced no similar effects on the CD spectrum of Cml, nor were comparable effects of NAG in evidence on the Cm reduced derivatives of ribonuclease, chymotrypsin, wheat germ agglutinin, or alpha-lactalbumin. The effect therefore appears specific between NAG and Cml. Conversion of the tryptophan residue at Position 62 of Cml to the oxindolealanyl derivative prevented these effects of NAG, and this residue may therefore participate in the interaction. During a 4-day incubation at room temperature, the analog preserved the CD spectrum of Cml as well as its concentration. This effect was nearly specific when compared with other Cm reduced proteins and with other carbohydrates. Only one, N-acetyl mannosamine, was effective in preserving the concentration of Cml, but not the CD spectrum. Since D-glucosamine was entirely without effect on either the CD spectrum of Cml or on its change during incubation, the acetyl group appears essential for the NAG-Cml interaction. The specificity between NAG and Cml is tentatively accounted for in terms of interactions with the primary structure, rather than with the remaining secondary structure.(ABSTRACT TRUNCATED AT 250 WORDS)
D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D002645:Chickens; D002942:Circular Dichroism; D004220:Disulfides; D009113:Muramidase / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D004220:Disulfides; D009113:Muramidase; D000117:Acetylglucosamine