Sunita Patel

Fluorescence Investigation of the Binding of Model PDT Drugs to Nonionic and Zwitterionic Surfactants

The binding of two model photodynamic therapy drugs, chlorin p6 and purpurin 18, with surfactants has been studied using steady‐state and time‐resolved fluorescence techniques. The surfactants used are amphiphilic nonionic surfactant (Tween 80 and Tween 40) and zwitterionic surfactant (HAPS). These have applications in drug delivery. The studies have been performed at pH 7 and 5 for chlorin p6 and at pH 7 for purpurin 18. The binding constants have been estimated from the change in fluorescence parameters and have been compared with those for Cremophor EL and human serum albumin. Chlorin p6 is found to bind to the surfactants to a greater extent at pH 5 than at pH 7. The same ionic species of chlorin p6 is found to exist at the maximum concentrations of the surfactants.

Competitive binding of Chlorin p6 and Dansyl-l-Proline to Sudlow’s site II of human serum albumin

The binding of chlorin p6, a model photosensitizer for photodynamic therapy (PDT), to the Sudlows site II of Human Serum Albumin (HSA) has been monitored by different spectroscopic methods. Displacement of Dansyl-l-Proline (DP) from its conjugate with HSA is manifested in the spectral shift and decrease in its fluorescence intensity as well as the emergence of component with lifetime of 2-3ns, which is characteristic of free DP. As DP is known to bind specifically to the Sudlows site II of human serum albumin, its displacement by chlorin p6 indicates the residence of the photosensitizer in the same site, in addition to Sudlows site I. The binding constants for Sudlows site II, determined by the stopped-flow technique, are found to be two orders of magnitude smaller than that for Sudlows site I.

A small tripeptide AFA undergoes two state cooperative conformational transitions: Implications for conformational biases in unfolded states

It is important to understand the conformational biases that are present in unfolded states to understand protein folding. In this context, it is surprising that even a short tripeptide like AFA samples folded/ordered conformation as demonstrated recently by NMR experiments of the peptide in aqueous solution at 280 K. In this paper, we present molecular dynamics simulation of the peptide in explicit water using OPLS-AA/L all-atom force field. The results are in overall agreement with NMR results and provide some further insights. The peptide samples turn and extended conformational forms corresponding to minima in free energy landscape. Frequent transitions between the minima are observed due to modest free energy barriers. The turn conformation seems to be stabilized by hydrophobic interactions and possibly by bridging water molecules between backbone donors and acceptors. Thus the peptide does not sample conformations randomly, but samples well defined conformations. The peptide served as a model for folding-unfolding equilibrium in the context of peptide folding. Further, implications for drug design are also discussed.

Mechanism of Initiation, Association, and Formation of Amyloid Fibrils Modeled with the N-Terminal Peptide Fragment, IKYLEFIS, of Myoglobin G-Helix

Some peptides and proteins undergo self-aggregation under certain conditions, leading to amyloid fibrils formation, which is related to many disease conditions. It is important to understand such amyloid fibrils formation to provide mechanistic detail that governs the process. A predominantly α-helical myoglobin has been reported recently to readily form amyloid fibrils at a higher temperature, similar to its G-helix segment. Here, we have investigated the mechanism of amyloid fibrils formation by performing multiple long molecular dynamics simulations (27 μs) on the N-terminal segment of the G-helix of myoglobin. These simulations resulted in the formation of a single-layered tetrameric β-sheet with mixed parallel and antiparallel β-strands and this is the most common event irrespective of many different starting structures. Formation of the single-layered tetrameric β-sheet takes place following three distinctive pathways. The process of fibril initiation is dependent on temperature. Further, this study provides mechanistic insights into the formation of multilayered fibrilar structure, which could be applicable to a wider variety of peptides or proteins to understand the amyloidogenesis.