The ternary complex of Mn2+, synthetic decapeptide DP1 (DEHGTAVMLK), and orthophosphate is a superb antioxidant

• 成果类型: Article
• 署名作者: Yang, Hao; Sharma, Ajay; Daly, Michael J.; Hoffman, Brian M.
• 署名单位: Northwestern University; Uniformed Services University of the Health Sciences - USA; Northwestern University
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-12756
• DOI: 10.1073/pnas.2417389121
• 发表日期: 2024-12-09
• 关键词: oxidative stress resistance; radiation-resistance; deinococcus-radiodurans; mn2+-peptide complex; escherichia-coli; gamma-radiation; protein damage; manganese; mitochondria; speciation

摘要

Mn2+ coordinated by orthophosphate (Pi), metabolites, or peptides acts as a superoxide dismutase (SOD), and these Mn antioxidant complexes are universally accumulated in extremely radiation- resistant cell types across the tree of life. This behavior prompted design of decapeptide DP1 (DEHGTAVMLK) as a Mn2+ ligand, and development of a highly potent Mn2+- antioxidant (MDP) containing [Pi] = 25 mM, and [DP1] = 3 mM, the ratio found in the radioresistant bacterium Deinococcus radiodurans, with [Mn2+] = 1 mM. MDP is an exceptional antioxidant, both in vitro and in vivo, and has reinvigorated the development of radiation- inactivated whole- cell vaccines. This study investigates the nature of the active Mn2+ complex in MDP. We measure the affinity of DP1 for the substitutionally labile Mn2+ ion using isothermal- titration calorimetry (ITC) and use changes in the Mn2+ solution EPR spectrum to determine affinities of Mn2+ for DP1 and for Pi, and to monitor Mn2+ ligation while titrated with the fixed Pi/DP1 ratio of MDP, 25/3, using ENDOR/ESEEM to characterize DP1 ligation to Mn2+. In parallel, 1 H NMR of DP1 was used to monitor binding interactions between Pi and DP1, and DP1 binding to the diamagnetic Ca2+. We report: i) DP1 forms an extremely weak, dynamic Mn2+ complex (Ka approximate to 40 M-1) ii) Mn2+ binds Pi much more strongly (Ka approximate to 390 M-1) as shown previously, but iii) DP1 and Pi jointly bind to Mn2+ in MDP to form a ternary Mn2+ (Pi) (DP1) complex with greater formation- constant than Pi alone (K a app approximate to 670 M-1). It is this ternary complex that is the superb antioxidant in MDP.