The copper-linked Escherichia coli AZY operon: Structure, metal binding, and a possible physiological role in copper delivery

Rose C. Hadley, Daniel Zhitnitsky, Nurit Livnat-Levanon, Gal Masrati, Elena Vigonsky, Jessica Rose, Nir Ben-Tal, Amy C. Rosenzweig, Oded Lewinson

نتاج البحث: نشر في مجلةمقالةمراجعة النظراء

ملخص

The Escherichia coli yobA-yebZ-yebY (AZY) operon encodes the proteins YobA, YebZ, and YebY. YobA and YebZ are homologs of the CopC periplasmic copper-binding protein and the CopD putative copper importer, respectively, whereas YebY belongs to the uncharacterized Domain of Unknown Function 2511 family. Despite numerous studies of E. coli copper homeostasis and the existence of the AZY operon in a range of bacteria, the operon's proteins and their functional roles have not been explored. In this study, we present the first biochemical and functional studies of the AZY proteins. Biochemical characterization and structural modeling indicate that YobA binds a single Cu2+ ion with high affinity. Bioinformatics analysis shows that YebY is widespread and encoded either in AZY operons or in other genetic contexts unrelated to copper homeostasis. We also determined the 1.8 Å resolution crystal structure of E. coli YebY, which closely resembles that of the lantibiotic self-resistance protein MlbQ. Two strictly conserved cysteine residues form a disulfide bond, consistent with the observed periplasmic localization of YebY. Upon treatment with reductants, YebY binds Cu+ and Cu2+ with low affinity, as demonstrated by metal-binding analysis and tryptophan fluorescence. Finally, genetic manipulations show that the AZY operon is not involved in copper tolerance or antioxidant defense. Instead, YebY and YobA are required for the activity of the copper-related NADH dehydrogenase II. These results are consistent with a potential role of the AZY operon in copper delivery to membrane proteins.

اللغة الأصليةالإنجليزيّة
رقم المقال101445
دوريةJournal of Biological Chemistry
مستوى الصوت298
رقم الإصدار1
المعرِّفات الرقمية للأشياء
حالة النشرنُشِر - 1 يناير 2022

All Science Journal Classification (ASJC) codes

  • !!Molecular Biology
  • !!Biochemistry
  • !!Cell Biology

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