Proline‐Rich Chaperones Are Compared Computationally and Experimentally for Their Abilities to Facilitate Recombinant Butyrylcholinesterase Tetramerization in CHO Cells. Issue 3 (17th November 2017)

• Record Type: Journal Article
• Authors: Wang, Qiong; Chen, Charles H.; Chung, Cheng‐yu; Priola, Joseph; Chu, Jeffrey H.; Tang, Juechun; Ulmschneider, Martin B.; Betenbaugh, Michael J.
• Is Part Of: Biotechnology journal. Volume 13:Issue 3(2018)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2017-11-17
• Subjects: butyrylcholinesterase tetramerization -- BChE -- proline‐rich chaperones -- molecular dynamic simulations -- Chinese hamster ovary; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.201700479
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract : Human butyrylcholinesterase (BChE), predominantly tetramers with a residence time of days, offers the potential to scavenge organophosphorus pesticides and chemical warfare agents. Efficient assembly of human BChE into tetramers requires an association with proline‐rich peptide chaperones. In this study, the incorporation of different proline‐rich peptide chaperones into BChE is investigated computationally and experimentally. First, the authors applied molecular dynamic (MD) simulations to interpret the interactions between proline‐rich chaperones with human BChE tetramer domains. The P24 chaperone which contains 24 prolines, promoted the association of BChE tetramer with a 74% simulated helicity of BChE subunits, whereas the control without chaperone and BChE with an 8‐proline chaperone (P8) complex exhibited 55.8 and 60.6% predicted helicity, respectively. The interaction of proline‐rich chaperones with BChE subunits (B‐P) provides a conduit to facilitate the interactions between BChE subunits (B‐B) of the complex, which is mainly attributed to hydrophobic interactions and hydrogen‐bond binding. Experimental assessment of these two proline‐rich chaperones plus a 14‐proline chaperone (P14) was performed and confirmed that P24 has superior capability to facilitate recombinant BChE (rBChE) tetramerization with >60% rBChE tetramer in P24‐transfected rBChE cells, whereas P14‐ and P8‐transfected rBChE cells had 44 and 33% rBChE tetramer, respectively. The rBChE control had 14% tetramer. Finally, we developed a stable rBChE tetramer expression system in CHO cells by enriching P24 expression in rBChE expressing cells. Overall, our simulations provided a design concept for identifying proline‐rich peptides that promote the rBChE tetramerization in CHO cells. Abstract : Efficient assembly of human butyrylcholin‐esterase (BChE) into tetramers requires its association with proline‐rich chaperones. In this study, the authors computationally and experientially assessed different proline‐rich chaperones (P8, P14, P24) for their abilities to facilitate recombinant BChE in vivo tetramerization in CHO cells. Consistent with computational simulation, P24 with longer proline length has superior capability to promote recombinant BChE tetramerization (>60%) than P8 and P14 chaperones in experimental study.