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06:58
Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling
Published on: October 7, 2021
In vitro translation and computational analyses of human profilin multimers
Michael Babich1, Lisa R P Foti, Linda Wong
1Department of Biomedical Sciences, University of Illinois College of Medicine, Rockford, Illinois, USA. michaelb@immvarx.com
Proceedings of the Western Pharmacology Society
|January 19, 2006
Summary
Human profilin proteins (P1 and P2) self-associate into stable multimers, forming disulfide bridges. This multimerization is crucial for their biological functions, including roles in cell signaling and allergies.
Area of Science:
- Biochemistry
- Molecular Biology
- Allergology
Background:
- Profilin is a ubiquitous G-actin and PIP2-binding protein known as a pan-allergen.
- Functional native human profilin multimers have been identified, suggesting roles in cell morphology, signaling, and allergies.
Purpose of the Study:
- To investigate the multimerization of human profilin I (P1) and II (P2) using cDNA.
- To analyze the structural basis of profilin self-association and its resistance to reducing agents.
Main Methods:
- In vitro transcription and radiolabeled translation of human P1 and P2 cDNAs.
- Autoradiography and immunoblotting of affinity column-purified proteins.
- Computer-based molecular modeling to analyze potential disulfide bonding and multimer structures.
Main Results:
- Profilin P1 and P2 cDNAs encode proteins that form stable multimers.
- Autoradiography revealed predominant 14.8, 30, and 58 kDa bands, indicating multimer formation.
- Molecular modeling suggested disulfide bridges, particularly between C16 and C127 in P1, forming a protected dimer structure.
Conclusions:
- Human profilin P1 and P2 proteins form stable, fastidious multimers.
- A structural model proposes protected disulfide bridges between profilin units, consistent with biochemical data.
- Profilin multimerization is a significant factor in its biological activities, including allergic responses.
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