STIM1-ORAI1 direct interaction cannot govern store-operated calcium entry (SOCE) in platelets
Introduction
Platelets play an essential role in hemostasis as they prevent blood loss upon vessel wall disruption
The growing complexity of platelet aggregation
S. P. Jackson
Blood. 2007, 109, 5087-5095
Calcium signaling in platelets
D. VARGA-SZABO, A. BRAUN, B. NIESWANDT
Journal of Thrombosis and Haemostasis. 2009, 7, 1057-1066
Calcium signalling: IP3 rises again… and again
C. Taylor, P. Thorn
Current Biology. 2001, 11, R352-R355
Voltage-gated calcium channels, calcium signaling, and channelopathies
E. S. Piedras-Rentería, C. F. Barrett, Y.-Q. Cao, and R. W. Tsien
New Comprehensive Biochemistry. 2007, 41, 127-166
Two main proteins contributing to SOCE are ER protein STIM1, that senses Ca2+ levels in intracellular stores
STIM Is a Ca2+ Sensor Essential for Ca2+-Store-Depletion-Triggered Ca2+ Influx
J. Liou, M. Kim, W. Do Heo, J. Jones, J. Myers, J. Ferrell, T. Meyer
Current Biology. 2005, 15, 1235-1241
The STIM1/Orai signaling machinery
M. Fahrner, I. Derler, I. Jardin, C. Romanin
Channels. 2013, 7, 330-343
Roles of Platelet STIM1 and Orai1 in Glycoprotein VI- and Thrombin-dependent Procoagulant Activity and Thrombus Formation
K. Gilio, R. van Kruchten, A. Braun, A. Berna-Erro, M. Feijge, D. Stegner, P. van der Meijden, M. Kuijpers, D. Varga-Szabo, J. Heemskerk, B. Nieswandt
Journal of Biological Chemistry. 2010, 285, 23629-23638
What is the mechanism of SOCE in platelets? While the crucial role of STIM1 and ORAI1 proteins is well established
Regulation of Platelet Function by Orai, STIM and TRP
A. Berna-Erro, I. Jardín, T. Smani, and J. A. Rosado
Calcium Entry Pathways in Non-excitable Cells. 2016, 898, 157-181
Multifaceted roles of STIM proteins
R. Hooper, E. Samakai, J. Kedra, J. Soboloff
Pflügers Archiv - European Journal of Physiology. 2013, 465, 1383-1396
The STIM1/Orai signaling machinery
M. Fahrner, I. Derler, I. Jardin, C. Romanin
Channels. 2013, 7, 330-343
More Than Just Simple Interaction between STIM and Orai Proteins: CRAC Channel Function Enabled by a Network of Interactions with Regulatory Proteins
S. Berlansky, C. Humer, M. Sallinger, I. Frischauf
International Journal of Molecular Sciences. None, 22, 471
Essential Role for the CRAC Activation Domain in Store-dependent Oligomerization of STIM1
E. Covington, M. Wu, R. Lewis
Molecular Biology of the Cell. 2010, 21, 1897-1907
STIM1 Clusters and Activates CRAC Channels via Direct Binding of a Cytosolic Domain to Orai1
C. Park, P. Hoover, F. Mullins, P. Bachhawat, E. Covington, S. Raunser, T. Walz, K. Garcia, R. Dolmetsch, R. Lewis
Cell. 2009, 136, 876-890
Single-molecule analysis of diffusion and trapping of STIM1 and Orai1 at endoplasmic reticulum–plasma membrane junctions
M. Wu, E. Covington, R. Lewis
Molecular Biology of the Cell. 2014, 25, 3672-3685
How strict is the correlation between STIM1 and Orai1 expression, puncta formation, and ICRAC activation?
T. Gwozdz, J. Dutko-Gwozdz, V. Zarayskiy, K. Peter, V. Bolotina
American Journal of Physiology-Cell Physiology. 2008, 295, C1133-C1140
Numbers count: How STIM and Orai stoichiometry affect store-operated calcium entry
M. Yen, R. Lewis
Cell Calcium. 2019, 79, 35-43
Besides direct STIM1-ORAI1 interaction there is an alternative SOCE mechanism. It requires intermediate diffusible messenger – calcium influx factor (CIF)
Orai, STIM1 and iPLA2β: a view from a different perspective
V. Bolotina
The Journal of Physiology. 2008, 586, 3035-3042
Novel Role for STIM1 as a Trigger for Calcium Influx Factor Production
P. Csutora, K. Peter, H. Kilic, K. Park, V. Zarayskiy, T. Gwozdz, V. Bolotina
Journal of Biological Chemistry. 2008, 283, 14524-14531
There are several mathematical models of SOCE
Systems biology insights into the meaning of the platelet's dual-receptor thrombin signaling
A. Sveshnikova, A. Balatskiy, A. Demianova, T. Shepelyuk, S. Shakhidzhanov, M. Balatskaya, A. Pichugin, F. Ataullakhanov, M. Panteleev
Journal of Thrombosis and Haemostasis. 2016, 14, 2045-2057
Systems Modeling of Ca2+ Homeostasis and Mobilization in Platelets Mediated by IP3 and Store-Operated Ca2+ Entry
A. Dolan, S. Diamond
Biophysical Journal. 2014, 106, 2049-2060
Reaction-diffusion model for STIM-ORAI interaction: The role of ROS and mutations
B. Schmidt, D. Alansary, I. Bogeski, B. Niemeyer, H. Rieger
Journal of Theoretical Biology. 2019, 470, 64-75
Reaction-diffusion model for STIM-ORAI interaction: The role of ROS and mutations
B. Schmidt, D. Alansary, I. Bogeski, B. Niemeyer, H. Rieger
Journal of Theoretical Biology. 2019, 470, 64-75
Models of Solute Aggregation Using Cellular Automata
L. Kier, C. Cheng, J. Nelson
Chemistry & Biodiversity. 2009, 6, 396-401
In this article we propose a lattice-based model of SOCE in platelets. We demonstrate that small surface concentration of STIM1 cannot provide sufficient stoichiometry for proper pore opening and Ca2+ influx.
Modelling approach
The SOCE model is defined by two discrete spatial lattices \( \mathcal{L} \): ER and PM membranes, a discrete state space \( \varepsilon \) and local rule-based dynamics.
The regular lattice \( \mathcal{L} \subset {\rm I\!R^2} \) consists of \( N \) nodes \( r_{i} \in \mathcal{L}, i=1, \ldots, N \). Every node has \( b=8 \) nearest neighbors. Each lattice node \( \ r \in \mathcal{L} \) is connected to its nearest neighbor by unit vectors \( c_{i}, i=1, \ldots, b \). The neighborhood is assumed to be Moore neighborhood. In addition, protein can be in a resting state, that is presented as \( c_0 \). As we consider two opposing lattices, that denote two membranes (ER and PM), additional unit vector \( c_9 \) is introduced in order to represent the occupation in the second lattice at same lattice cell. The parameter \( K=b+1 \) defines the possible node movement choices.
The state space is defined through the occupation numbers \( s_{j} \in\{0,1\}, j=0, \ldots, K \). These occupation numbers represent the presence \( s_{j}=1 \) or absence \( s_{j}=0 \) of a protein in the channel \( c_j \) within some node. Then, the configuration of a node is given by the state vector $$s=\left(s_{1}, \ldots, s_{K}\right) \in \varepsilon=\{0,1\}^{K}$$
The node by itself has properties. In our model STIM1(ORAI1) can be in two states – free, bound. Additionally, STIM1 can be a part of a cluster. This fact is denoted by the parameter \( a \in\{0,1,2\} \), where \( a=0 \) represents free protein, \( a=1 \) – bound, \(a=2 \) – in cluster (for STIM1). If a protein is in bound state it can no longer diffuse along the membrane (Fig. 1).
A new lattice configuration is created according to a local rule that determines the new state of each node in terms of the current states of the node and the nodes in its neighborhood. In order to determine a new lattice configuration, the local rule is applied sequentially at every node \( r \) of the lattice.
The algorithm, by which STIM1-ORAI1 interaction was modelled is presented in the following block-diagram (Fig.2):
Note that we assume that close location of proteins is not sufficient for clustering and they need to “overlap” in order to form a cluster. Therefore, we consider clustering as a process that occurs when one protein tends to diffuse “into” another protein, rather than cluster due to simple proximity.
Initial state of each membrane is produced by randomly placing nodes on 2-dimentional lattice. The number of nodes for each lattice is equal to the number of proteins (STIM1 or ORAI1).
Model parameters were assessed manually in order to match experimental data. Specifically, STIM1-ORAI1 probability was estimated according to known characteristic times and maximal number of STIM1-ORAI1 complexes and STIM1 clustering according to known STIM1:ORAI1 stoichiometry within one puncta.
Results
Evaluation of diffusion coefficient
Our modelling approach for diffusion is similar to modelling Brownian motion using a random walk. However, length of the step for each iteration is fixed and is equal to 1 lattice cell. This is due to the fact that when simulating the Brownian motion of particles in water, the collision length of a particle is much larger than the size of the particle itself due to the liquid state of aggregation of the medium
Applied Biophysics: A Molecular Approach for Physical Scientists
T. A. Waigh
Applied Biophysics. 2007, None, None
Single-molecule analysis of diffusion and trapping of STIM1 and Orai1 at endoplasmic reticulum–plasma membrane junctions
M. Wu, E. Covington, R. Lewis
Molecular Biology of the Cell. 2014, 25, 3672-3685
STIM1-ORAI1 direct interaction in HEK cells
In order to investigate the mechanism of SOCE in platelets, we first applied our model on HEK cells. Such approach was chosen based on two reasons: 1) HEK cells have well known geometric parameters and protein counts, including typical PM-ER junction areas, 2) STIM1-ORAI1 direct interaction is mostly investigated on this type of cells and so there is plenty of experimental data, on which model can be evaluated. The parameters for HEK cells were taken from
Reaction-diffusion model for STIM-ORAI interaction: The role of ROS and mutations
B. Schmidt, D. Alansary, I. Bogeski, B. Niemeyer, H. Rieger
Journal of Theoretical Biology. 2019, 470, 64-75
Typical system state for ER membrane on \( t=0 \mathrm{~ms}, 10 \mathrm{~ms} \text { and } 100 \mathrm{~ms} \) is shown in Fig. 4A. The model parameters were estimated to match rapid STIM1-ORAI1 puncta formation (
Reaction-diffusion model for STIM-ORAI interaction: The role of ROS and mutations
B. Schmidt, D. Alansary, I. Bogeski, B. Niemeyer, H. Rieger
Journal of Theoretical Biology. 2019, 470, 64-75
Dynamic Coupling of the Putative Coiled-coil Domain of ORAI1 with STIM1 Mediates ORAI1 Channel Activation
M. Muik, I. Frischauf, I. Derler, M. Fahrner, J. Bergsmann, P. Eder, R. Schindl, C. Hesch, B. Polzinger, R. Fritsch, H. Kahr, J. Madl, H. Gruber, K. Groschner, C. Romanin
Journal of Biological Chemistry. 2008, 283, 8014-8022
SOCE modelling in platelets
After estimating parameters for STIM1-ORAI1 interaction we applied our model on platelets. We used same probabilities for clustering and binding, but changed the initial number of proteins on each membrane. It appears, that such low STIM1 concentration cannot provide normal stoichiometry. Typical system states are presented in Fig.5A. It also seems that characteristic time of f STIM1-ORAI1 puncta formation is enhanced (Fig. 5B).
Discussion
In the present work we developed a lattice-based model of STIM1-ORAI1 interaction. The model was able to adequately describe protein coupling dynamics in HEK cells, which was in accordance to experimental data and results of different modelling approach. Our data also revealed that in HEK cells crosslinking of ORAI1 pores by STIM1 takes place, that according to
Cross-linking of Orai1 channels by STIM proteins
Y. Zhou, R. Nwokonko, X. Cai, N. Loktionova, R. Abdulqadir, P. Xin, B. Niemeyer, Y. Wang, M. Trebak, D. Gill
Proceedings of the National Academy of Sciences. 2018, 115, E3398-E3407
On the other side, in platelets not only crosslinking does not occur, but neither needed stoichiometry is achieved. This is due to small surface STIM1 concentration, even if all STIM1 proteins are considered to be fully on ER, although they are presented both on PM and ER.
There are several ways to interpret the obtained data. Firstly, we may assume that direct interaction does not play significant role in platelets. Then, as STIM1 and ORAI1 are known to mediate platelet function, we consider different mechanism of SOCE. iPLA2 pathway of SOCE is known to be present in platelets and may be responsible for SOCE instead of direct coupling model. However, it was shown by Harper et. al.
Is calcium-independent phospholipase A2required for store-operated calcium entry in human platelets?
M. HARPER, S. SAGE
Journal of Thrombosis and Haemostasis. 2008, 6, 1819-1821
Another possible mechanism of SOCE still accounts direct interaction of STIM1 and ORAI1, but with addition of TRPC channels
STIM1, Orai1 and hTRPC1 are important for thrombin- and ADP-induced aggregation in human platelets
C. Galán, H. Zbidi, A. Bartegi, G. Salido, J. Rosado
Archives of Biochemistry and Biophysics. 2009, 490, 137-144
TRPC Channels in the SOCE Scenario
J. Lopez, I. Jardin, J. Sanchez-Collado, G. Salido, T. Smani, J. Rosado
Cells. 2020, 9, 126
Author Contributions
Funding
Conflicts of Interest
References of this article:
The growing complexity of platelet aggregation
S. P. Jackson
Blood. 2007, 109, 5087-5095
Calcium signaling in platelets
D. VARGA-SZABO, A. BRAUN, B. NIESWANDT
Journal of Thrombosis and Haemostasis. 2009, 7, 1057-1066
Calcium signalling: IP3 rises again… and again
C. Taylor, P. Thorn
Current Biology. 2001, 11, R352-R355
Voltage-gated calcium channels, calcium signaling, and channelopathies
E. S. Piedras-Rentería, C. F. Barrett, Y.-Q. Cao, and R. W. Tsien
New Comprehensive Biochemistry. 2007, 41, 127-166
STIM Is a Ca2+ Sensor Essential for Ca2+-Store-Depletion-Triggered Ca2+ Influx
J. Liou, M. Kim, W. Do Heo, J. Jones, J. Myers, J. Ferrell, T. Meyer
Current Biology. 2005, 15, 1235-1241
The STIM1/Orai signaling machinery
M. Fahrner, I. Derler, I. Jardin, C. Romanin
Channels. 2013, 7, 330-343
Roles of Platelet STIM1 and Orai1 in Glycoprotein VI- and Thrombin-dependent Procoagulant Activity and Thrombus Formation
K. Gilio, R. van Kruchten, A. Braun, A. Berna-Erro, M. Feijge, D. Stegner, P. van der Meijden, M. Kuijpers, D. Varga-Szabo, J. Heemskerk, B. Nieswandt
Journal of Biological Chemistry. 2010, 285, 23629-23638
Regulation of Platelet Function by Orai, STIM and TRP
A. Berna-Erro, I. Jardín, T. Smani, and J. A. Rosado
Calcium Entry Pathways in Non-excitable Cells. 2016, 898, 157-181
Multifaceted roles of STIM proteins
R. Hooper, E. Samakai, J. Kedra, J. Soboloff
Pflügers Archiv - European Journal of Physiology. 2013, 465, 1383-1396
More Than Just Simple Interaction between STIM and Orai Proteins: CRAC Channel Function Enabled by a Network of Interactions with Regulatory Proteins
S. Berlansky, C. Humer, M. Sallinger, I. Frischauf
International Journal of Molecular Sciences. , 22, 471
Essential Role for the CRAC Activation Domain in Store-dependent Oligomerization of STIM1
E. Covington, M. Wu, R. Lewis
Molecular Biology of the Cell. 2010, 21, 1897-1907
STIM1 Clusters and Activates CRAC Channels via Direct Binding of a Cytosolic Domain to Orai1
C. Park, P. Hoover, F. Mullins, P. Bachhawat, E. Covington, S. Raunser, T. Walz, K. Garcia, R. Dolmetsch, R. Lewis
Cell. 2009, 136, 876-890
Single-molecule analysis of diffusion and trapping of STIM1 and Orai1 at endoplasmic reticulum–plasma membrane junctions
M. Wu, E. Covington, R. Lewis
Molecular Biology of the Cell. 2014, 25, 3672-3685
How strict is the correlation between STIM1 and Orai1 expression, puncta formation, and ICRAC activation?
T. Gwozdz, J. Dutko-Gwozdz, V. Zarayskiy, K. Peter, V. Bolotina
American Journal of Physiology-Cell Physiology. 2008, 295, C1133-C1140
Numbers count: How STIM and Orai stoichiometry affect store-operated calcium entry
M. Yen, R. Lewis
Cell Calcium. 2019, 79, 35-43
Orai, STIM1 and iPLA2β: a view from a different perspective
V. Bolotina
The Journal of Physiology. 2008, 586, 3035-3042
Novel Role for STIM1 as a Trigger for Calcium Influx Factor Production
P. Csutora, K. Peter, H. Kilic, K. Park, V. Zarayskiy, T. Gwozdz, V. Bolotina
Journal of Biological Chemistry. 2008, 283, 14524-14531
Systems biology insights into the meaning of the platelet's dual-receptor thrombin signaling
A. Sveshnikova, A. Balatskiy, A. Demianova, T. Shepelyuk, S. Shakhidzhanov, M. Balatskaya, A. Pichugin, F. Ataullakhanov, M. Panteleev
Journal of Thrombosis and Haemostasis. 2016, 14, 2045-2057
Systems Modeling of Ca2+ Homeostasis and Mobilization in Platelets Mediated by IP3 and Store-Operated Ca2+ Entry
A. Dolan, S. Diamond
Biophysical Journal. 2014, 106, 2049-2060
Reaction-diffusion model for STIM-ORAI interaction: The role of ROS and mutations
B. Schmidt, D. Alansary, I. Bogeski, B. Niemeyer, H. Rieger
Journal of Theoretical Biology. 2019, 470, 64-75
Models of Solute Aggregation Using Cellular Automata
L. Kier, C. Cheng, J. Nelson
Chemistry & Biodiversity. 2009, 6, 396-401
Applied Biophysics: A Molecular Approach for Physical Scientists
T. A. Waigh
Applied Biophysics. 2007, ,
Dynamic Coupling of the Putative Coiled-coil Domain of ORAI1 with STIM1 Mediates ORAI1 Channel Activation
M. Muik, I. Frischauf, I. Derler, M. Fahrner, J. Bergsmann, P. Eder, R. Schindl, C. Hesch, B. Polzinger, R. Fritsch, H. Kahr, J. Madl, H. Gruber, K. Groschner, C. Romanin
Journal of Biological Chemistry. 2008, 283, 8014-8022
Cross-linking of Orai1 channels by STIM proteins
Y. Zhou, R. Nwokonko, X. Cai, N. Loktionova, R. Abdulqadir, P. Xin, B. Niemeyer, Y. Wang, M. Trebak, D. Gill
Proceedings of the National Academy of Sciences. 2018, 115, E3398-E3407
Is calcium-independent phospholipase A2required for store-operated calcium entry in human platelets?
M. HARPER, S. SAGE
Journal of Thrombosis and Haemostasis. 2008, 6, 1819-1821
STIM1, Orai1 and hTRPC1 are important for thrombin- and ADP-induced aggregation in human platelets
C. Galán, H. Zbidi, A. Bartegi, G. Salido, J. Rosado
Archives of Biochemistry and Biophysics. 2009, 490, 137-144
TRPC Channels in the SOCE Scenario
J. Lopez, I. Jardin, J. Sanchez-Collado, G. Salido, T. Smani, J. Rosado
Cells. 2020, 9, 126