2-Methyl-2,4-pentanediol (MPD) boosts as detergent-substitute the performance of ß-barrel hybrid catalyst for phenylacetylene polymerization

Abstract

Covering hydrophobic regions with stabilization agents to solubilize purified transmembrane proteins is crucial for their application in aqueous media. The small molecule 2-methyl-2,4-pentanediol (MPD) was used to stabilize the transmembrane protein Ferric hydroxamate uptake protein component A (FhuA) utilized as host for the construction of a rhodium-based biohybrid catalyst. Unlike commonly used detergents such as sodium dodecyl sulfate or polyethylene polyethyleneglycol, MPD does not form micelles in solution. Molecular dynamics simulations revealed the effect and position of stabilizing MPD molecules. The advantage of the amphiphilic MPD over micelle-forming detergents is demonstrated in the polymerization of phenylacetylene, showing a ten-fold increase in yield and increased molecular weights.

Unravelling the refolding of membrane proteins, one of the most challenging therapeutic targets

“…Remarkably, our method has successfully been applied to several peptides and proteins featuring diverse structures and properties [9][10][11][12][13][14]. Indeed, several monomeric β-barrel MPs with a variable number of transmembrane (TM) strands have been refolded: PagP (8TM), a VDAC (19TM), FhuA (22TM).…”

“…Oligomeric MPs have also been recovered: the heptameric hemolysin (14TM) and the trimeric Omp2a (16TM). Aside, our seminal technique was recently used for diverse applications, taking advantage of its refolding, solubilizing and stabilizing properties [10,[15][16][17]. Moreover, crystal structures of both the globular hen-egg lysozyme [18] and the membrane protein PagP [19] have clearly established the preservation of the native state in a SDS-MPD mixture.…”

Exploring and Adapting the Molecular Selectivity of Artificial Metalloenzymes

“…Looking at rhodium-based ArMs, several studies have proven their capacity using a variety of different protein scaffolds to perform reactions such as enantio-and chemoselective hydrogenation, 92-98 NADP + reduction, 99 hydroformylation, 100 [4+2] annulation, [101][102] and phenylacetylene polymerization. [103][104][105][106][107][108] Additionally, carbene transfer reactions (alkene cyclopropanation, [109][110][111][112] N−H insertion, 109 S−H insertion, 109 Si−H insertion, 109,111 C−H insertion 112 ) have also been extensively investigated. In one study of significance, the biocompatibility of a nitrobindin-based ArM was evaluated on microbial cell surfaces.…”

Current Applications of Artificial Metalloenzymes and Future Developments

“…Another ArM was built up by covalent anchoring of the same rhodium(cod) complex to an engineered form of Fhu A [93]. [Note: Description of FhuA and its engineered form ∆CVF will be given below].…”