"No gene hits!" error when using Daphnia pulex pathway library with UniProt or KEGG protein IDs

Hello,

I am trying to perform a Joint Pathway Analysis integrating metabolomics and proteomics data using the Daphnia pulex pathway library in MetaboAnalyst 6.0. However, I consistently receive a “No gene hits!” error when I provide my protein list, regardless of whether I use UniProt IDs or KEGG IDs specific to D. pulex.

However, if I replace the D. pulex UniProt IDs with the UniProt IDs of their Homo sapiens orthologs and switch the pathway library to Homo sapiens, the analysis runs without any issue. I think that problem is specific to the D. pulex pathway library not recognizing the protein identifiers.

Steps performed (leading to the error):

1. Opened Pathway Analysis in MetaboAnalyst 6.0.

2. Selected Joint Pathway Analysis

3. For metabolomics, I first attempted Untargeted metabolomics input; after encountering the error, I switched to Targeted analysis using compound names as identifiers.

4. Uploaded the protein list using:

- D. pulex-specific UniProt IDs (first attempt), then

- D. pulex KEGG IDs (second attempt).

5. Selected Daphnia pulex as the pathway library.

6. Clicked to proceed — error message received: “No gene hits!”

**Working with Homo sapiens orthologs:**

- Same workflow as above, but using Homo sapiens orthologs UniProt IDs and selecting the Homo sapiens pathway library → analysis completes successfully.

- I am attaching screenshots of both analyses (targeted and untargeted)

- Two showing the successful run with the **Homo sapiens** pathway library.

- Two documenting the **“No gene hits!”** error with the **Daphnia pulex** pathway library.

Thank you for your help!

Please provide your actual input (i.e. not screenshot of the input)?

Thank you for your response, my input is:

#CompoundList
D-glucose
Pyruvate
DG

#Uniprot_Dpulex
E9HF20
E9HLR0
E9G8H5
E9GS42
E9GZ78

#Uniprot_Hsapiens
P49721
P49720
O96008
P28072
Q9BVA1

The same list of compounds was used for the analyses of both Daphnia pulex and the orthologous proteins of Homo sapiens.

Hi Cyn,
Thanks for sharing these details. I can reproduce this issue. It will be fixed in a few days and I will update you here once it is ready to use.
Bests,
Zhiqiang

It is fixed. Please try again after server update this weekend. Thank you for the note

Hi! Thank you so much for your response and help. I can confirm that the problem is resolved and the analysis ran without any issues. :slightly_smiling_face:

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