Here are POSGOST67's posts about Dr. Urano's u

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There is one way to do this. You can make smaller and smaller mutants of recombinant MANF and use an assay where you apply those MANF mutants to neurons, and have a reporter that lights up when the neurons are dividing, you can compare full length MANF to the fragments and find the portion that is responsible for activating the receptor.

That will give you an amino acid sequence that binds to the receptor, then you can use that small MANF sequence as your "small molecule" or use it to narrow down your choices for what the actual receptor .There is one way to do this. You can make smaller and smaller mutants of recombinant MANF and use an assay where you apply those MANF mutants to neurons, and have a reporter that lights up when the neurons are dividing, you can compare full length MANF to the fragments and find the portion that is responsible for activating the receptor.

That will give you an amino acid sequence that binds to the receptor, then you can use that small MANF sequence as your "small molecule" or use it to narrow down your choices for what the actual receptor are.
What's the receptor for MANF?The receptor for extracellular MANF has never been found. There was one manuscript a while back that hinted at KDEL receptors, (receptors that bind to proteins that contain that 4 amino acid sequence K-D-E-L), but this has never been verified.
I don't think he will find a small molecule that induces MANF production that is not cytotoxic. Basically, it would be very difficult to find a molecule that selectively induces MANF production.

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.I don't think he will find a small molecule that induces MANF production that is not cytotoxic. Basically, it would be very difficult to find a molecule that selectively induces MANF production.



Funding is scarce now. He is protecting his lab. A small molecule can be purchased or produced for a fraction of what the cost of producing or purchasing MANF. If it doesn't work out, oh well, you didn't waste that much money. If it works out, you saved your lab a ton of money.
Small molecules are cheaper to manufacture and more stable. Whole recombinant proteins are expensive to manufacture and generally less stable.

That said, MANF is produced when cells undergo an endoplasmic reticulum stress response, which means that the small molecules that will induce it cause some cytotoxicity (cell stress/cell death).

Unless he has looked at the promoter region of the MANF gene and found some specific transcriptional pathways that induce MANF, I think that small molecule inducers of MANF may be bad news

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