Structural Exploration on Palmitoyltransferase DHHC3 from Homo sapiens

Area of Science:

• Biochemistry
• Structural Biology
• Molecular Biology

Background:

• DHHC3 is a palmitoyltransferase enzyme crucial for protein modification.
• DHHC3 is a potential target for cancer therapeutics, but its structure and mechanism are unknown.
• Sequence diversity complicates structural studies compared to known DHHC homologues.

Purpose of the Study:

• To express and purify human DHHC3 (hDHHC3) and a truncated version.
• To identify interacting proteins that facilitate hDHHC3 crystallization.
• To perform initial structural exploration of hDHHC3 using cryo-electron microscopy (cryo-EM).

Main Methods:

• Protein expression and purification of hDHHC3 and its N-terminal domain (NTD) deleted mutant.
• Mass spectrometry and co-immunoprecipitation to identify interacting proteins (LAMTOR1).
• Cryo-electron microscopy (cryo-EM) for structural analysis of an inactive hDHHC3 mutant.

Main Results:

• Successfully expressed and purified hDHHC3 and a truncated variant.
• Identified LAMTOR1 as an interacting protein aiding in crystallization.
• Obtained preliminary cryo-EM data showing a typical membrane protein sideview for an inactive hDHHC3 mutant.

Conclusions:

• The study provides a foundation for future structural determination of hDHHC3.
• Understanding DHHC3 structure is critical for developing targeted cancer therapies.
• The identified interacting protein LAMTOR1 may play a role in DHHC3 function or regulation.