Structure-driven development of a biomimetic rare earth artificial metalloprotein

• 成果类型: Article
• 署名作者: Thompson, Peter J.; Boggs, David G.; Wilson, Charles A.; Bruchs, Austin T.; Velidandla, Uditha; Bridwell-Rabb, Jennifer; Olshansky, Lisa
• 署名单位: University of Illinois System; University of Illinois Urbana-Champaign; University of Michigan System; University of Michigan; University of Illinois System; University of Illinois Urbana-Champaign
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-11350
• DOI: 10.1073/pnas.2405836121
• 发表日期: 2024-08-13
• 关键词: methanol dehydrogenase; active-transport; protein; binding; MODEL; lanthanides; ions

摘要

The 2011 discovery of the first rare earth-dependent enzyme in methylotrophic Methylobacterium extorquens AM1 prompted intensive research toward understanding the unique chemistry at play in these systems. This enzyme, an alcohol dehydrogenase (ADH), features a La3+ ion closely associated with redox- active coenzyme pyrroloquinoline quinone (PQQ) and is structurally homologous to the Ca2+- dependent ADH from the same organstructurally characterized to 1.46- & Aring; resolution by X- ray diffraction. This crystal structure reveals a Lys residue hydrogen- bonded to PQQ at the site analogously occupied by a Lewis acidic cation in ADH. Accordingly, we prepared K142A- and K142D- PqqT variants to assess the relevance of this site toward metal binding. Isothermal titration calorimetry experiments and produces spectral signatures consistent with those of ADH enzymes. These spectral zyme primed for future mechanistic, biocatalytic, and biosensing applications.