TY - JOUR
T1 - Chemical Synthesis of TFF3 Reveals Novel Mechanistic Insights and a Gut-Stable Metabolite
AU - Braga Emidio, Nayara
AU - Meli, Rajeshwari
AU - Tran, Hue N T
AU - Baik, Hayeon
AU - Morisset-Lopez, Séverine
AU - Elliott, Alysha G
AU - Blaskovich, Mark A T
AU - Spiller, Sabrina
AU - Beck-Sickinger, Annette G
AU - Schroeder, Christina I
AU - Muttenthaler, Markus
N1 - Publisher Copyright:
© 2021 The Authors. Published by American Chemical Society.
PY - 2021/7/8
Y1 - 2021/7/8
N2 - TFF3 regulates essential gastro- and neuroprotective functions, but its molecular mode of action remains poorly understood. Synthetic intractability and lack of reliable bioassays and validated receptors are bottlenecks for mechanistic and structure-activity relationship studies. Here, we report the chemical synthesis of TFF3 and its homodimer via native chemical ligation followed by oxidative folding. Correct folding was confirmed by NMR and circular dichroism, and TFF3 and its homodimer were not cytotoxic or hemolytic. TFF3, its homodimer, and the trefoil domain (TFF310-50) were susceptible to gastrointestinal degradation, revealing a gut-stable metabolite (TFF37-54; t1/2 > 24 h) that retained its trefoil structure and antiapoptotic bioactivity. We tried to validate the putative TFF3 receptors CXCR4 and LINGO2, but neither TFF3 nor its homodimer displayed any activity up to 10 μM. The discovery of a gut-stable bioactive metabolite and reliable synthetic accessibility to TFF3 and its analogues are cornerstones for future molecular probe development and structure-activity relationship studies.
AB - TFF3 regulates essential gastro- and neuroprotective functions, but its molecular mode of action remains poorly understood. Synthetic intractability and lack of reliable bioassays and validated receptors are bottlenecks for mechanistic and structure-activity relationship studies. Here, we report the chemical synthesis of TFF3 and its homodimer via native chemical ligation followed by oxidative folding. Correct folding was confirmed by NMR and circular dichroism, and TFF3 and its homodimer were not cytotoxic or hemolytic. TFF3, its homodimer, and the trefoil domain (TFF310-50) were susceptible to gastrointestinal degradation, revealing a gut-stable metabolite (TFF37-54; t1/2 > 24 h) that retained its trefoil structure and antiapoptotic bioactivity. We tried to validate the putative TFF3 receptors CXCR4 and LINGO2, but neither TFF3 nor its homodimer displayed any activity up to 10 μM. The discovery of a gut-stable bioactive metabolite and reliable synthetic accessibility to TFF3 and its analogues are cornerstones for future molecular probe development and structure-activity relationship studies.
KW - Biophysical Phenomena
KW - HEK293 Cells
KW - Humans
KW - Molecular Structure
KW - Oxidation-Reduction
KW - Protein Folding
KW - Structure-Activity Relationship
KW - Trefoil Factor-3/chemical synthesis
UR - http://www.scopus.com/inward/record.url?scp=85110139988&partnerID=8YFLogxK
U2 - 10.1021/acs.jmedchem.1c00767
DO - 10.1021/acs.jmedchem.1c00767
M3 - Journal article
C2 - 34142550
SN - 0022-2623
VL - 64
SP - 9484
EP - 9495
JO - Journal of Medicinal Chemistry
JF - Journal of Medicinal Chemistry
IS - 13
ER -