In Silico Characterization of Calcineurin from Pathogenic Obligate Intracellular Trypanosomatids: Potential New Biological Roles
Abstract
:1. Introduction
2. Materials and Methods
2.1. Database Inspection
2.2. Obtaining Consensus Sequences and Multiple Alignment of CaN
2.3. Evaluation of Conserved Domains, Determination of Physicochemical Parameters, and Hydrophobicity Profiles
2.4. In Silico Identification of Protein–Protein Interactions of CaN Regulatory Subunits
2.5. Prediction of the Subcellular Localization of CaN in Pathogenic Intracellular Trypanosomatids
2.6. Prediction of Entry to the Non-Classical Secretory Pathway of CaN in Pathogenic Intracellular Trypanosomatids
2.7. Prediction of Cleavage by Calpains in CaN Catalytic Subunits of Pathogenic Intracellular Trypanosomatids
2.8. Prediction of Phosphorylation Sites in the CaM-Binding Domain (CaM-BD) of CaN Catalytic Subunits from Leishmania Spp.
3. Results
3.1. The CaN Subunits of Obligate Intracellular Trypanosomatids Possess Diverse, Partially Conserved Domain Architecture and Potential Calpain Cleavage Sites
3.2. The CaM-Binding Domain in Leishmania Spp. Has Greater Potential to Be Regulated by Phosphorylation Than Its Human Ortholog
3.3. CaN Regulatory Subunits of Obligate Intracellular Trypanosomatids Differ in Their Calcium-Binding Domains from Their Human Counterpart and Myristoylation Potential, but Preserve Some Canonical EF-Loops and the Docking Site for Immunophilin–Immunosuppressive Drug Complexes
3.4. CaN Regulatory and Catalytic Subunits of Obligate Intracellular and Human Trypanosomatids Would Share the Subcellular Distribution Pattern, with Some Exceptions
3.5. CaN Regulatory Subunits of Obligate Intracellular Trypanosomatids Interact Only with Their Catalytic Monomer and Related Immunophilins
3.6. CaN Regulatory and Catalytic Subunits of Obligate Intracellular Trypanosomatids Have a Differential Potential to Be Secreted by the Non-Classical Pathway
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Protein | #aa | MW (Da) | pI |
---|---|---|---|
HsCaNAα | 521 | 58,687.85 | 5.58 |
TcCaNA1 | 382 | 43,244.64 | 4.83 |
TcCaNA2 | 392 | 44,619.50 | 8.13 |
LsppCaNA1 | 545 | 59,747.65 | 5.00 |
LsppCaNA1_var | 503 | 55,559.36 | 5.61 |
LsppCaNA2 | 407 | 45,138.00 | 6.37 |
TcCaNB | 176 | 19,458.18 | 4.98 |
LsppCaNB | 176 | 19,785.54 | 4.60 |
HsCaNAα | #x | Context | Score | Kinase | Answer |
---|---|---|---|---|---|
# Sequence | 21 S | ARVFSVLRE | 0.974 | unsp | YES |
# Sequence | 27 S | LREES---- | 0.556 | PKA | YES |
LsppCaNA1 and LsppCaNA1var | #x | Context | Score | Kinase | Answer |
# Sequence | 17 S | MGRLSRMFH | 0.956 | unsp | YES |
# Sequence | 17 S | MGRLSRMFH | 0.725 | PKA | YES |
# Sequence | 17 S | MGRLSRMFH | 0.556 | PKC | YES |
# Sequence | 22 T | RMFHTLCEG | 0.766 | Unsp | YES |
# Sequence | 22 T | RMFHTLCEG | 0.517 | CKII | YES |
A. WoLF PSORT of Catalytic Subunits of CaN | ||||||
---|---|---|---|---|---|---|
Site | HsCaNAα | TcCaNA1 | TcCaNA2 | LsppCaNA1 | LsppCaNA1_var | LsppCaNA2 |
extr | 3 | |||||
plas | 1 | |||||
cyto | 21.5 | 12 | 8.5 | 13.5 | 14.5 | 9.5 |
cysk | 6 | |||||
E.R. | 3 | 1 | ||||
golg | 1 | |||||
mito | 6 | 3 | 1 | 2 | 9 | |
pero | 3 | 2 | 5 | 3 | 1 | 2 |
vacu | 1 | |||||
nucl | 4 | 4 | 9.5 | 3 | 9.5 | 3.5 |
cyto_nucl | 13.5 | 9 | 10.5 | 10 | 13.5 | 7 |
B. WoLF PSORT of Regulatory Subunits of CaN | ||||||
Site | HsCaNB-1 | TcCaNB | LsppCaNB | |||
extr | 1 | 1 | ||||
plas | 2 | |||||
cyto | 25 | 5.5 | 9 | |||
cysk | 1 | 6 | ||||
mito | 1 | |||||
pero | 3 | 3 | 2 | |||
nucl | 2 | 9.5 | 15 | |||
Cyto_nucl | 14.5 | 8 | 14 |
Name | NN-Score 1 | Odds | Weighted by Prior | Warning |
---|---|---|---|---|
HsCaNAα | 0.441 | 0.861 | 0.002 | - |
HsCaNB-1 | 0.548 | 1.252 | 0.003 | - |
TcCaNA1 | 0.682 | 1.863 | 0.004 | - |
TcCaNA2 | 0.442 | 0.858 | 0.002 | - |
TcCaNB | 0.637 | 1.758 | 0.004 | - |
LsppCaNA1 | 0.528 | 1.094 | 0.002 | - |
LsppCaNA1_var | 0.477 | 0.926 | 0.002 | - |
LsppCaNA2 | 0.718 | 2.165 | 0.004 | - |
LsppCaNB | 0.382 | 0.709 | 0.001 | - |
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Orrego, P.R.; Serrano-Rodríguez, M.; Cortez, M.; Araya, J.E. In Silico Characterization of Calcineurin from Pathogenic Obligate Intracellular Trypanosomatids: Potential New Biological Roles. Biomolecules 2021, 11, 1322. https://doi.org/10.3390/biom11091322
Orrego PR, Serrano-Rodríguez M, Cortez M, Araya JE. In Silico Characterization of Calcineurin from Pathogenic Obligate Intracellular Trypanosomatids: Potential New Biological Roles. Biomolecules. 2021; 11(9):1322. https://doi.org/10.3390/biom11091322
Chicago/Turabian StyleOrrego, Patricio R., Mayela Serrano-Rodríguez, Mauro Cortez, and Jorge E. Araya. 2021. "In Silico Characterization of Calcineurin from Pathogenic Obligate Intracellular Trypanosomatids: Potential New Biological Roles" Biomolecules 11, no. 9: 1322. https://doi.org/10.3390/biom11091322