Amino acid function relates to its embedded protein microenvironment: A study on disulfide-bridged cystine

ABSTRACT In our previous study, we have shown that the microenvironments around conserved amino acids are also conserved in protein families (Bandyopadhyay and Mehler, Proteins 2008; 72:646–659). In this study, we have hypothesized that amino acids perform similar functions when embedded in a certain type of protein microenvironment. We have tested this hypothesis on the microenvironments around disulfide‐bridged cysteines from high‐resolution protein crystal structures. Although such cystines mainly play structural role in proteins, in certain enzymes they participate in catalysis and redox reactions. We have performed and report a functional annotation of enzymatically active cystines to their respective microenvironments. Three protein microenvironment clusters were identified: (i) buried‐hydrophobic, (ii) exposed‐hydrophilic, and (iii) buried‐hydrophilic. The buried‐hydrophobic cluster encompasses a small group of 22 redox‐active cystines, mostly in alpha‐helical conformations in a –C‐x‐x‐C‐ motif from the Oxido‐reductase enzyme class. All these cystines have high strain energy and near identical microenvironments. Most of the active cystines in hydrolase enzyme class belong to buried hydrophilic microenvironment cluster. In total there are 34 half‐cystines detected in buried hydrophilic cluster from hydrolases, as a part of enzyme active site. Even within the buried hydrophilic cluster, there is clear separation of active half‐cystines between surface exposed part of the protein and protein interior. Half‐cystines toward the surface exposed region are higher in number compared to those in protein interior. Apart from cystines at the active sites of the enzymes, many more half‐cystines were detected in buried hydrophilic cluster those are part of the microenvironment of enzyme active sites. However, no active half‐cystines were detected in extremely hydrophilic microenvironment cluster, that is, exposed hydrophilic cluster, indicating that total exposure of cystine toward the solvent is not favored for enzymatic reactions. Although half‐cystines in exposed‐hydrophilic clusters occasionally stabilize enzyme active sites, as a part of their microenvironments. Analysis performed in this work revealed that cystines as a part of active sites in specific enzyme families or folds share very similar protein microenvironment regions, despite of their dissimilarity in protein sequences and position specific sequence conservations. Proteins 2016; 84:1576–1589. ©