Allosteric regulation is a fundamental mechanism employed by cells to control critical processes such as signal transduction, catalysis, and gene regulation [1-3]. While studies of allosteric regulation have often focused on thermodynamic aspects, there has been an increasing realization of the role of conformational dynamics [4-5].  Recent advances in NMR [6], cryo electron microscopy [7-8] and biophysical techniques that enabled detailed investigations of large protein systems at atomic resolution have fueled the resurgence of computational and theoretical studies of allosteric regulation, leading to new conceptual outlooks of this long-standing biological phenomenon [9]. High-pressure NMR experiments can be used to detect of low-lying excited functional states, providing another tool to investigate the dynamic energy landscapes [10]. Simulation-based computational approaches allow substantive comparative studies of allosteric networks of regulation and are increasingly being combined with NMR and cryo-EM investigations [11-12]. The ability to measure accurate distances and kinetics with Single-molecule FRET (smFRET)  has led to its emergence as an important tool for mapping biomolecular heterogeneities and for measuring structural dynamics over a wide range of timescales [13-15]. Despite the established view  that  complex  protein systems  and  regulatory complexes often function as dynamic and versatile allosteric machines, the characterization of  hidden and rare protein functional states,  allosteric conformational transformations and allosteric pathways    is still surprisingly limited,  calling for  the integration of  novel structural, biophysical and computational approaches to address these challenges.

The overarching idea of this meeting is to build on the great success and overwhelming positive feedback of our previous CECAM Workshop on “Multiscale simulations of allosteric regulatory mechanisms…” (2018) and   CECAM Workshop  “Quantifying Protein Dynamics and Allosteric regulation in the cell with emerging technologies: From Cryo-EM and NMR to Networks and Machine Learning” ( 2021).

Our symposium  will assemble  top researchers in computational structural biology, drug design, crystallography, NMR, cryo-EM, single molecule spectroscopy  together with computational biologists, theoreticians, computer scientists and machine learning experts. Among confirmed speakers are members of the USA National Academy of Sciences  and Royal Society. The meeting will feature  many prominent computational biologists and top crystallographers, NMR experts and leaders in cryo-EM technology and FRET approaches as well as leaders in AI and Machine Learning.

We bring together an exciting  group of experts in  diverse areas : from computational  and structural biology,  theory and simulations to biophysics, to artificial intelligence, machine learning and  drug discovery.

The format of our meeting will include not only lectures and oral presentations but also a series of round table discussions,  focused group sessions, and poster presentations.

We are planning to cover a number of topics including  but not limited to :

a) Theoretical and computational models of allosteric regulation on different time scales:  accelerated MD simulations, Markov State models, non-equilibrium simulation methods.

b)  Experiment-informed modeling of allosteric biomolecular assemblies, network models, dynamic network flows, NMR-based simulation approaches, systems-biology simulations of regulation in the cellular environment.

b) New methods and developments in the non-equilibrium simulation methods for  modeling  of allosteric ensembles and pathways

c) Latest developments in  structural characterization of  allosteric molecular events and  hidden functional states important for allosteric function using cryo-EM, NMR, smFRET spectroscopy and integrative computational biophysics  approaches.

d)  Computer simulation methods and experimental NMR, smFRET tools   for  unveiling the invisible aspects of protein ‘life’ including the determination  of hidden protein states.

e) Integration of smFRET  tools with advanced sampling and non-equilibrium simulation methods for studies of slow conformational dynamics  and  allosteric transformations in protein systems

f) Open questions and challenges in the field, particularly opportunities for integration of  NMR, smFRET and advanced  computational sampling approaches.

The main objectives of the workshop are:

1. Focus on  the latest developments in  structural characterization of  allosteric molecular events and  hidden functional states important for allosteric function using cryo-EM, NMR, smFRET spectroscopy and integrative computational biophysics  approaches.
2. Discuss progress and identify integrative tools   for  unveiling the invisible aspects of protein ‘life’ including the determination  of hidden protein states.
3. Analyze and discuss new methods and developments in the non-equilibrium simulation methods for  modeling  of allosteric ensembles and pathways.
4. Focus on  approaches for  integration of smFRET  tools with advanced sampling and non-equilibrium simulation methods for studies of slow conformational dynamics  and  allosteric transformations in protein systems.
5. Develop a perspective on the progress and role of emerging technologies in quantifying dynamics  and kinetics of allosteric  events and  detecting hidden rare functional states  (high-pressure NMR,  cryo-EM. smFRET, experiment-guided biophysical modeling, AI and machine learning as enabling tools for integration of the theory and experiment).
6. Discuss open issues and challenges in the field, particularly opportunities for integration of  NMR, smFRET and advanced  computational sampling approaches
7. Bring together  cryo-EM. NMR, smFRET and computational communities to develop strategic views on allosteric phenomena in molecular biology.
8. Discuss Open Science,  Shared Infrastructure and  Data Exchange between cryo-EM, NMR, smFRET and computational communities to develop strategic views on allosteric phenomena in molecular biology.
9. Open exchange and discussion about new developments and current status in the field.
10. Provide opportunities and engage  students and early-career researchers to discuss their projects in a poster session and contributed talks. 
11. Address gender inequality in science by promoting participation of  women and minorities.
12.  Promote networking between students, early-career and more experienced researchers.

New Focused Topics  for Talks and Round-Table  Discussions: 

• COVID-19 Research : New Era for Structural and Computational Biology
• Advances in Artificial Intelligence and Impact on  Structural Biology, Biomedical Sciences and Integrative Biophysical Approaches
• AlphaFold Present and Future : From Structures to Dynamics, Mechanisms and Engineering
• ChatGPT  OpenAI :  Future of Science, Education and Publishing