Idioma:

Lanthanide-dependent alcohol dehydrogenases require an essential aspartate residue for metal coordination and enzymatic function

Good, Nathan M. ; Fellner, Matthias ; Demirer, Kemal ; Hu, Jian ; Hausinger, Robert P. ; Martinez-Gomez, N. Cecilia

The Journal of biological chemistry, 2020-06, Vol.295 (24), p.8272-8284 [Periódico revisado por pares]

United States: Elsevier Inc

Texto completo disponível

Citações Citado por

Enviar para

Título:
Lanthanide-dependent alcohol dehydrogenases require an essential aspartate residue for metal coordination and enzymatic function
Autor: Good, Nathan M. ; Fellner, Matthias ; Demirer, Kemal ; Hu, Jian ; Hausinger, Robert P. ; Martinez-Gomez, N. Cecilia
Assuntos: alcohol dehydrogenase (ADH) ; Alcohol Oxidoreductases - chemistry ; Alcohol Oxidoreductases - metabolism ; Amino Acid Substitution ; aspartate (aspartic acid) ; Aspartic Acid - metabolism ; Bacterial Proteins - chemistry ; Bacterial Proteins - metabolism ; BASIC BIOLOGICAL SCIENCES ; Biocatalysis - drug effects ; Calcium - pharmacology ; cofactor coordination ; crystallography ; Crystallography, X-Ray ; ethanol dehydrogenase ; ExaF ; INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY ; lanthanide ; Lanthanoid Series Elements - pharmacology ; metalloprotein ; Methanol - pharmacology ; methanol dehydrogenase ; Methylobacterium extorquens - drug effects ; Methylobacterium extorquens - enzymology ; Methylobacterium extorquens - growth & development ; one-carbon metabolism ; Oxidation-Reduction ; Protein Structure and Folding ; pyrroloquinoline quinone ; Structure-Activity Relationship ; XoxF
É parte de: The Journal of biological chemistry, 2020-06, Vol.295 (24), p.8272-8284
Notas: 1750003; CHE-1516126
National Science Foundation (NSF)
Edited by Ruma Banerjee
These authors contributed equally to this work.
Descrição: The lanthanide elements (Ln3+), those with atomic numbers 57–63 (excluding promethium, Pm3+), form a cofactor complex with pyrroloquinoline quinone (PQQ) in bacterial XoxF methanol dehydrogenases (MDHs) and ExaF ethanol dehydrogenases (EDHs), expanding the range of biological elements and opening novel areas of metabolism and ecology. Other MDHs, known as MxaFIs, are related in sequence and structure to these proteins, yet they instead possess a Ca2+-PQQ cofactor. An important missing piece of the Ln3+ puzzle is defining what features distinguish enzymes that use Ln3+-PQQ cofactors from those that do not. Here, using XoxF1 MDH from the model methylotrophic bacterium Methylorubrum extorquens AM1, we investigated the functional importance of a proposed lanthanide-coordinating aspartate residue. We report two crystal structures of XoxF1, one with and another without PQQ, both with La3+ bound in the active-site region and coordinated by Asp320. Using constructs to produce either recombinant XoxF1 or its D320A variant, we show that Asp320 is needed for in vivo catalytic function, in vitro activity, and La3+ coordination. XoxF1 and XoxF1 D320A, when produced in the absence of La3+, coordinated Ca2+ but exhibited little or no catalytic activity. We also generated the parallel substitution in ExaF to produce ExaF D319S and found that this variant loses the capacity for efficient ethanol oxidation with La3+. These results provide evidence that a Ln3+-coordinating aspartate is essential for the enzymatic functions of XoxF MDHs and ExaF EDHs, supporting the notion that sequences of these enzymes, and the genes that encode them, are markers for Ln3+ metabolism.
Editor: United States: Elsevier Inc
Idioma: Inglês

Links

Voltar para lista de resultados

Realização: Logos de Redes Sociais:

Lanthanide-dependent alcohol dehydrogenases require an essential aspartate residue for metal coordination and enzymatic function

Good, Nathan M. ; Fellner, Matthias ; Demirer, Kemal ; Hu, Jian ; Hausinger, Robert P. ; Martinez-Gomez, N. Cecilia

United States: Elsevier Inc

Buscando em bases de dados remotas. Favor aguardar.