Reference to the native isopeptide bond structure allows comparison of the
non-native linkages with the atomic connectivity of the native isopeptide
linkage. The table colour coding is given as follows: orange,
protein/peptide of interest (POI) present in a reactant or in the non-native
linkage; green, Ub (or polyUb) present in a reactant or in the non-native
linkage (independent of the POI); red, small molecule reactant which forms a
component of the non-native linkage; black, chemical auxiliary present in a
reactant but not in the final conjugate conjugate; blue, blocking
functionality that permits iterative chain assembly. The main reagents for a
specific ligation procedure are provided but not all small molecule reagents
necessary have been specified. Atom labels, X, denotes the native amino acid
position whereas “X”, denotes a non-native amino acid or
mutation introduced at position X (for example, 76 and ‘76’,
referring to Gly-76 of Ub). Mutations of the native Ub amino acid sequence
at positions not directly relevant to the isopeptide bond are given in
parentheses within the reactant chemical structures.