| The R&R consensus sequence is a sequence of amino acids that have been found through sequence alignment studies to be relevant in chitin binding thus forming a chitin binding domain. Previous studies indicated that mutations of Y128 and F136 (tyrosine and phenylalanine) or mutations of T95 and D97 (threonine and aspartic acid) eliminated chitin binding (Rebers and Willis 2001). This suggested that these particular amino acids were crucial in the protein structure thus allowing chitin to bind. Threonine at position 95 was later hypothesized by Hamodrakas et al. (2005) to play a crucial role in the binding cleft of the RR-1 cuticular protein HCCP12 which was known to bind chitin. In order to achieve this goal, molecular models of the native protein and other models of the mutated proteins were created. A series of programs including AutoDockTools were used for the docking and interpretation of binding interactions between a six-unit macromolecule of chitin and the proposed models of the AGCP2B chitin binding protein from Anopheles gambiae. The results of the study showed that the intermolecular energies were all relatively close for the mutations made. Analysis of an original mutation attempted by Rebers and Willis (2001), (GST+65YF) yielded similar results suggesting that the docking analysis was ineffective in determining the dynamics of the chitin binding between the Anopheles gambiae protein and chitin. |
