Recently a technique based on the interaction between adhesion proteins extracted from Streptococcus pyogenes, known as SpyRing, has been widely used to improve the thermal resilience of enzymes, the assembly of biostructures, cancer cell recognition and other fields. In SpyRing, the two termini of the target enzyme are respectively linked to the peptide SpyTag and its protein partner SpyCatcher. SpyTag spontaneously reacts with SpyCatcher to form an isopeptide bond, with which the target enzyme forms a close ring structure. It was believed that the covalent cyclization of protein skeleton caused by SpyRing reduces the conformational entropy of biological structure and improves its rigidity, thus improving the thermal resilience of the target enzyme. However, the effects of SpyTag/ SpyCatcher interaction with this enzyme are poorly understood, and their regulation of enzyme properties remains unclear. Here, for simplicity, we took the single domain enzyme lichenase from Bacillus subtilis 168 as an example, studied the interface interactions in the SpyRing system by molecular dynamics simulations, and examined the effects of the changes of electrostatic interaction and van der Waals interaction on the thermal resilience of target enzyme. The simulations showed that the interface between SpyTag/SpyCatcher and lichenase is different from that found by geometric matching method and highlighted key mutations that affect the intensity of interactions at the interface and might have effect on the thermal resilience of the enzyme. Our calculations provided new insights into the rational designs in the SpyRing.
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