pedH

UniProt ID: Q88JH0
Organism: Pseudomonas putida KT2440
Review Status: DRAFT
Aliases:
qedH-II PP_2679
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Gene Description

Lanthanide-dependent pyrroloquinoline quinone (PQQ)-dependent alcohol dehydrogenase that catalyzes periplasmic oxidation of diverse aliphatic and aromatic alcohols. PedH requires usable trivalent lanthanides for catalysis and is the metal-conditioned counterpart of calcium-dependent PedE. PedS2/PedR2 signaling and an unresolved positive input favor pedH expression when lanthanides are available; its physiological cytochrome c electron-transfer partner in KT2440 has not been directly established.

Proposed New Ontology Terms

lanthanide ion binding

Definition: Binding to a lanthanide ion, including lanthanum (La3+), cerium (Ce3+), praseodymium (Pr3+), neodymium (Nd3+), and samarium (Sm3+). Lanthanides are f-block elements with unique chemical properties distinct from transition metals.

Justification: Many bacterial enzymes specifically require lanthanides rather than calcium or other metals. Current GO only has generic "metal ion binding" which fails to capture this specificity.

lanthanide-dependent alcohol dehydrogenase activity

Definition: Catalysis of the oxidation of alcohols to aldehydes or ketones with the concomitant reduction of an electron acceptor, requiring lanthanide ions for activity.

Justification: Distinguishes lanthanide-dependent enzymes from calcium-dependent PQQ dehydrogenases, which is functionally and evolutionarily significant.

Existing Annotations Review

GO Term Evidence Action Reason
GO:0005509 calcium ion binding
IEA
GO_REF:0000120
REMOVE
Summary: Incorrect annotation. PedH is lanthanide-dependent, not calcium-dependent. This annotation likely arose from sequence similarity to other PQQ dehydrogenases. PedH requires La3+, Ce3+, Pr3+, Nd3+, Sm3+ etc. for activity, but not Ca2+.
GO:0016020 membrane
IEA
GO_REF:0000002
REMOVE
Summary: Too vague and potentially misleading. PedH is a periplasmic protein, not membrane-associated. While it has a signal peptide for periplasmic targeting, it is a soluble enzyme in the periplasm, not membrane-bound.
GO:0016614 oxidoreductase activity, acting on CH-OH group of donors
IEA
GO_REF:0000002
KEEP AS NON CORE
Summary: Accurate but too general. This correctly describes the broad enzymatic activity but lacks specificity about the PQQ-dependence and cytochrome c as electron acceptor. The more specific term GO:0052934 is present and preferred.
GO:0030288 outer membrane-bounded periplasmic space
IEA
GO_REF:0000002
REMOVE
Summary: Less accurate than GO:0042597. While PedH is in the periplasm, this term implies specific association with the outer membrane boundary. Bioinformatics analysis confirms PedH is a soluble enzyme (no TM regions in mature protein) that functions throughout the periplasmic space, not specifically at membrane interfaces. The hydrophobic region (aa 6-27) is the signal peptide, cleaved upon export.
Proposed replacements: periplasmic space
GO:0016491 oxidoreductase activity
IEA
GO_REF:0000120
REMOVE
Summary: Too general and uninformative. While technically correct, this term provides no specific functional information. The more specific terms GO:0052934 and GO:0016614 are present and much more informative.
GO:0020037 heme binding
IEA
GO_REF:0000117
REMOVE
Summary: Incorrect annotation. PedH is a PQQ quinoprotein rather than a heme-containing enzyme. A cytochrome c is the assigned reaction acceptor, not a prosthetic group bound by PedH.
GO:0042597 periplasmic space
IEA
GO_REF:0000120
ACCEPT
Summary: Correct and well-supported. PedH has a signal peptide (aa 1-25) directing export to periplasm. Bioinformatics analysis confirms no transmembrane regions in mature protein - it's a soluble enzyme functioning throughout the periplasmic space. Like all PQQ-ADHs (ExaA, PedE, MxaF, XoxF), PedH is freely diffusible in periplasm where it oxidizes alcohols and transfers electrons to cytochrome c.
Supporting Evidence:
file:PSEPK/pedH/pedH-deep-research-falcon.md
PedH is explicitly described as part of a **periplasmic oxidation system** (together with PedE) that supports growth on alcoholic substrates
GO:0046872 metal ion binding
IEA
GO_REF:0000043
ACCEPT
Summary: Correct but too general. PedH specifically binds lanthanide ions (La3+, Ce3+, Pr3+, Nd3+, Sm3+) which are essential for activity. This is NOT calcium-dependent like other PQQ dehydrogenases. GO lacks a specific lanthanide binding term which would be more accurate. Falcon deep research confirms lanthanide-exclusive activity and attributes the metal selectivity to a conserved active-site Asp (vs Ser in the Ca2+-dependent paralog PedE).
Supporting Evidence:
file:PSEPK/pedH/pedH-deep-research-falcon.md
PedH is catalytically active **only in the presence of trivalent lanthanides (Ln3+)** and is inactive with several tested heavy lanthanides/other trivalent metals.
file:PSEPK/pedH/pedH-deep-research-falcon.md
PedH contains a **conserved active-site Asp** (contrasting with a Ser in PedE) that is associated with **Ln3+ coordination** in lanthanide-dependent quinoprotein dehydrogenases.
GO:0052934 alcohol dehydrogenase (cytochrome c) activity
IEA
GO_REF:0000120
ACCEPT
Summary: Specific EC-derived annotation for PedH's core alcohol-oxidation chemistry. The purified-enzyme assays establish lanthanide-dependent alcohol dehydrogenase activity, but they used PMS/DCPIP rather than directly identifying the physiological KT2440 cytochrome c partner.
Supporting Evidence:
file:PSEPK/pedH/pedH-deep-research-falcon.md
PedH belongs to a family of periplasmic **quinoprotein** alcohol dehydrogenases that use the redox cofactor **PQQ** to catalyze oxidation of alcohols (and in some cases aldehydes).
file:PSEPK/pedH/pedH-deep-research-falcon.md
It oxidizes a **broad range** of alcohol substrates, including **linear and aromatic primary and secondary alcohols**, and can show activity with some aldehydes
GO:0070968 pyrroloquinoline quinone binding
IEA
GO_REF:0000117
ACCEPT
Summary: Perfect annotation. PedH absolutely requires PQQ as a cofactor (1:1 stoichiometry) for all catalytic activity. This is well-supported by crystal structure data showing specific PQQ binding sites. Core molecular function. Falcon deep research classifies PedH within the periplasmic PQQ-dependent quinoprotein alcohol dehydrogenase family.
Supporting Evidence:
file:PSEPK/pedH/pedH-deep-research-falcon.md
PedH belongs to a family of periplasmic **quinoprotein** alcohol dehydrogenases that use the redox cofactor **PQQ** to catalyze oxidation of alcohols (and in some cases aldehydes).
GO:0006066 alcohol metabolic process
IEA NEW
Summary: PedH catalyzes the periplasmic oxidation of a broad range of alcohols including linear aliphatic alcohols, aromatic alcohols, and secondary alcohols.
Reason: This biological process term captures PedH's primary function in alcohol catabolism, which is essential for growth on various volatile organic compounds.
Supporting Evidence:
PMID:28655819
efficient growth of cells with various naturally occurring alcoholic VOCs relies on the functional production of the lanthanide-dependent ADH PedH
file:PSEPK/pedH/pedH-deep-research.md
catalyzes the periplasmic oxidation of a broad range of alcohols including linear aliphatic alcohols, aromatic alcohols such as 2-phenylethanol, secondary alcohols
GO:0042537 benzene-containing compound metabolic process
IEA NEW
Summary: PedH is central to the 2-phenylethanol degradation pathway and catalyzes the oxidation of aromatic alcohols including 2-phenylethanol to phenylacetaldehyde.
Reason: This biological process term captures PedH's specific role in aromatic compound catabolism, particularly in the degradation of benzene-containing alcohols.
Supporting Evidence:
PMID:18177365
Another catabolic route (PedS(1)R(1)ABCS(2)R(2)DEFGHI) is needed for the uptake of 2-phenylethanol and for its oxidation to phenylacetic acid via phenylacetaldehyde.
file:PSEPK/pedH/pedH-deep-research.md
PedH is integral to aromatic alcohol catabolism and detoxification in Pseudomonas putida. It is part of the 2-phenylethanol degradation (Ped) pathway
file:PSEPK/pedH/pedH-deep-research-falcon.md
a **Ξ”pedH** strain fails to grow on 2-phenylethanol under conditions where the REE switch represses reliance on PedE
GO:0010468 regulation of gene expression
IMP
PMID:28655819
Functional Role of Lanthanides in Enzymatic Activity and Tra...
NEW
Summary: PedH deletion changes pedE and pedH promoter output, establishing a non-core regulatory phenotype while leaving the molecular mechanism unresolved.
Reason: A BP annotation does not imply that PedH binds DNA or has a transcription-regulator MF. Retain the genetic feedback phenotype without adding such a molecular function or placing it in the catalytic core.
Supporting Evidence:
PMID:28655819
PedH not only has a catalytic function but is also involved in the transcriptional regulation of pedE and pedH, most likely acting as a sensory module
PMID:30158283
positive-feedback function of PedH in the presence of Ln3+ ions

Core Functions

Lanthanide-dependent alcohol dehydrogenase activity in periplasm for aromatic and aliphatic alcohol metabolism

Supporting Evidence:
  • PMID:28655819
    efficient growth of cells with various naturally occurring alcoholic VOCs relies on the functional production of the lanthanide-dependent ADH PedH
  • PMID:18177365
    conversion of 2-phenylethanol into phenylacetaldehyde and onward to phenylacetic acid
  • file:PSEPK/pedH/pedH-deep-research.md
    catalyzes the periplasmic oxidation of a broad range of alcohols including linear aliphatic alcohols, aromatic alcohols such as 2-phenylethanol, secondary alcohols
  • file:PSEPK/pedH/pedH-deep-research-falcon.md
    PedH is best understood as a **periplasmic, lanthanide-activated PQQ alcohol dehydrogenase** that broadens KT2440’s ability to oxidize diverse alcohols
  • file:PSEPK/pedH/pedH-deep-research-falcon.md
    It oxidizes a **broad range** of alcohol substrates, including **linear and aromatic primary and secondary alcohols**, and can show activity with some aldehydes

Pyrroloquinoline quinone cofactor binding for catalytic activity

Supporting Evidence:
  • PMID:28655819
    PedH (PP_2679) exhibits enzyme activity on a range of substrates similar to that of its Ca2+-dependent counterpart PedE (PP_2674), including linear and aromatic primary and secondary alcohols, as well as aldehydes, but only in the presence of lanthanide ions
  • file:PSEPK/pedH/pedH-deep-research.md
    Its primary structure contains a large eight-bladed Ξ²-propeller domain that harbors the PQQ prosthetic group
  • file:PSEPK/pedH/pedH-deep-research-falcon.md
    PedH belongs to a family of periplasmic **quinoprotein** alcohol dehydrogenases that use the redox cofactor **PQQ** to catalyze oxidation of alcohols (and in some cases aldehydes).

Lanthanide metal ion binding for enzymatic activation (requires La3+, Ce3+, Pr3+, Nd3+, or Sm3+ - not calcium)

Molecular Function:
metal ion binding
Supporting Evidence:
  • PMID:28655819
    PedH exhibits enzymatic activity only in the presence of lanthanides
  • PMID:30158283
    the transcription of the genes encoding the Ca2+-utilizing enzyme PedE and the Ln3+-utilizing enzyme PedH are inversely regulated
  • file:PSEPK/pedH/pedH-deep-research.md
    A critical active-site residue substitution (Asp in PedH vs Ser in PedE) enables PedH to coordinate lanthanides and confers its lanthanide reliance
  • file:PSEPK/pedH/pedH-deep-research-falcon.md
    PedH contains a **conserved active-site Asp** (contrasting with a Ser in PedE) that is associated with **Ln3+ coordination** in lanthanide-dependent quinoprotein dehydrogenases.
  • file:PSEPK/pedH/pedH-deep-research-falcon.md
    PedH is catalytically active **only in the presence of trivalent lanthanides (Ln3+)** and is inactive with several tested heavy lanthanides/other trivalent metals.

References

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Suggested Questions for Experts

Q: What structural features of PedH enable selective binding of lanthanides over more abundant calcium ions?

Q: How does lanthanide availability in the environment regulate the expression and activity of PedH versus calcium-dependent alcohol dehydrogenases?

Q: What is the electronic basis for the enhanced catalytic efficiency of lanthanide-PQQ complexes compared to calcium-PQQ?

Suggested Experiments

Experiment: X-ray absorption spectroscopy to characterize lanthanide coordination geometry and electronic structure in the active site

Experiment: Competition assays with various lanthanides to determine binding affinity hierarchy and catalytic efficiency

Experiment: Environmental metagenomics to correlate pedH gene abundance with soil lanthanide concentrations

Deep Research

Asta

(pedH-deep-research-asta.md)

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Falcon

(pedH-deep-research-falcon.md)

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OpenScientist

(pedH-deep-research-openscientist.md)

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Deep Research Report: pedH (PSEPK)

(pedH-deep-research.md)

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πŸ“š Additional Documentation

Notes

(pedH-notes.md)

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Bioinformatics Results

(RESULTS.md)

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Bioreason Rl Predictions

(pedH-bioreason-rl-predictions.md)

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Bioreason Rl Review

(pedH-bioreason-rl-review.md)

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