Optical biosensor investigation of interactions of biomembrane and water‐soluble cytochromes P450 and their redox partners with covalently immobilized phosphatidylethanolamine layers

Abstract

A phospholipid-containing biochip was created by covalently immobilizing phospholipids on the optical biosensor's aminosilane cuvette and employed to monitor the interactions of the membrane and water-soluble proteins in cytochrome P450-containing monooxygenase systems with planary layers of dilauroylphosphatidylethanolamine (DLPE) and distearoylphosphatidylethanolamine (DSPE), differing in acyl chain length. It was shown that the full-length membrane proteins-cytochrome P4502B4 (d-2B4), cytochrome b5 (d-b5) and NADPH-cytochrome P450 reductase (d-Fp)-readily incorporated into the phospholipids. The incorporation was largely due to hydrophobic interactions of membranous protein fragments with the phospholipid layer. However, electrostatic forces were also but not always involved in the incorporation process. They promoted d-Fp incorporation but had no effect on d-b5 incorporation. In low ionic strength buffer, no incorporation of these two proteins into the DSPE lipid layer was observable. Incorporation of d-b5 into the DLPE layer was abruptly increased at temperatures exceeding phospholipid phase transition point. Incorporation of d-2B4 was dependent on its aggregation state and decreased with increasing protein aggregability. Water-soluble proteins either would not interact with the phospholipid layer (adrenodoxin) or would bind to the layer at the cost of only electrostatic (albumin) or both electrostatic and hydrophobic (P450cam) interactions.

Direct electrochemistry of human, bovine and porcine cytochrome P450c17

“…During the past decade, direct electrochemistry has been obtained at edge-plane PGE without redox mediators (Kazlauskaite et al 1996), in lipid films (Ivanov et al 2001, Zhang et al 1997, using sol-gels (Iwuoha et al 2000), in colloidal clay (Lei et al 2000) and with surface-confined promoters (Aguey-Zinsou et al 2003, Lvov et al 1998, Munge et al 2003. Numerous other redox studies have examined P450s in solution.…”

Nanotechnologies in proteomics

“…Of all biosensor types, particular attention should be paid to noncontact OBs that have found an ever-widening application in biology and medicine [54][55][56][57][58][59][60][61][62][63][64][65][66][67][68][69]. They are used in the real-time analyses of protein -protein interactions including the interactions between antigen/antibody, oligonucleotide/ oligonucleotide, protein/lipid, hormone/receptor and, also, drug/receptor -with the latter interactions being widely used for drug design and drug testing.…”

A flexible and miniaturized hair dye based photodetector via chemiluminescence pathway

“…Optical chemical sensors possess high sensitivity, easy operation, high accuracy, and wide detection capacity. (Ratterman et al, 2014;Salama et al, 2007;Hatakeyama et al, 2009;Wang et al, 2015;Yamamoto et al, 2013;Ruano et al, 2000;Tsafack et al, 2000;Ivanov et al, 2001;Askari et al, 2002;Song et al, 2005) In particular, luminescence-based optical chemical sensors exhibit certain advantages, including a low detection limit, insensitivity to electrical disturbance, and a wide linear working range. (Delaney et al, 2011;Borisov et al, 2008).…”