Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces
Abstract Our previous study on the binding activity between Cel5H and clay minerals showed highest binding efficiency among other cellulase enzymes cloned. Here, based on previous studies, we hypothesized that the positive amino acids on the surface of Cel5H protein may play an important role in bin...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
SpringerOpen
2021-11-01
|
| Series: | Applied Microscopy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s42649-021-00066-7 |
| _version_ | 1830359628813172736 |
|---|---|
| author | Renukaradhya K. Math Nagakumar Bharatham Palaksha K. Javaregowda Han Dae Yun |
| author_facet | Renukaradhya K. Math Nagakumar Bharatham Palaksha K. Javaregowda Han Dae Yun |
| author_sort | Renukaradhya K. Math |
| collection | DOAJ |
| description | Abstract Our previous study on the binding activity between Cel5H and clay minerals showed highest binding efficiency among other cellulase enzymes cloned. Here, based on previous studies, we hypothesized that the positive amino acids on the surface of Cel5H protein may play an important role in binding to clay surfaces. To examine this, protein sequences of Bacillus licheniformis Cel5H (BlCel5H) and Paenibacillus polymyxa Cel5A (PpCel5A) were analyzed and then selected amino acids were mutated. These mutated proteins were investigated for binding activity and force measurement via atomic force microscopy (AFM). A total of seven amino acids which are only present in BlCel5H but not in PpCel5A were selected for mutational studies and the positive residues which are present in both were omitted. Of the seven selected surface lysine residues, only three mutants K196A(M2), K54A(M3) and K157T(M4) showed 12%, 7% and 8% less clay mineral binding ability, respectively compared with wild-type. The probable reason why other mutants did not show altered binding efficiency might be due to relative location of amino acids on the protein surface. Meanwhile, measurement of adhesion forces on mica sheets showed a well-defined maximum at 69 ± 19 pN for wild-type, 58 ± 19 pN for M2, 53 ± 19 pN for M3, and 49 ± 19 pN for M4 proteins. Hence, our results demonstrated that relative location of surface amino acids of Cel5H protein especially positive charged amino acids are important in the process of clay mineral-protein binding interaction through electrostatic exchange of charges. |
| first_indexed | 2024-12-20T03:03:04Z |
| format | Article |
| id | doaj.art-a99cb17f71c14186865bf020012d35cc |
| institution | Directory Open Access Journal |
| issn | 2287-4445 |
| language | English |
| last_indexed | 2024-12-20T03:03:04Z |
| publishDate | 2021-11-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | Applied Microscopy |
| spelling | doaj.art-a99cb17f71c14186865bf020012d35cc2022-12-21T19:55:41ZengSpringerOpenApplied Microscopy2287-44452021-11-0151111010.1186/s42649-021-00066-7Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forcesRenukaradhya K. Math0Nagakumar Bharatham1Palaksha K. Javaregowda2Han Dae Yun3SDM Research Institute for Biomedical Sciences, 5th Floor, Manjushree Building, SDM College of Medical Sciences & Hospital Campus, Shri Dharmasthala Manjunatheshwara UniversityThe University of Trans-Disciplinary Health Sciences and Technology (TDU)SDM Research Institute for Biomedical Sciences, 5th Floor, Manjushree Building, SDM College of Medical Sciences & Hospital Campus, Shri Dharmasthala Manjunatheshwara UniversityDivision of Applied Life Sciences, Gyeongsang National UniversityAbstract Our previous study on the binding activity between Cel5H and clay minerals showed highest binding efficiency among other cellulase enzymes cloned. Here, based on previous studies, we hypothesized that the positive amino acids on the surface of Cel5H protein may play an important role in binding to clay surfaces. To examine this, protein sequences of Bacillus licheniformis Cel5H (BlCel5H) and Paenibacillus polymyxa Cel5A (PpCel5A) were analyzed and then selected amino acids were mutated. These mutated proteins were investigated for binding activity and force measurement via atomic force microscopy (AFM). A total of seven amino acids which are only present in BlCel5H but not in PpCel5A were selected for mutational studies and the positive residues which are present in both were omitted. Of the seven selected surface lysine residues, only three mutants K196A(M2), K54A(M3) and K157T(M4) showed 12%, 7% and 8% less clay mineral binding ability, respectively compared with wild-type. The probable reason why other mutants did not show altered binding efficiency might be due to relative location of amino acids on the protein surface. Meanwhile, measurement of adhesion forces on mica sheets showed a well-defined maximum at 69 ± 19 pN for wild-type, 58 ± 19 pN for M2, 53 ± 19 pN for M3, and 49 ± 19 pN for M4 proteins. Hence, our results demonstrated that relative location of surface amino acids of Cel5H protein especially positive charged amino acids are important in the process of clay mineral-protein binding interaction through electrostatic exchange of charges.https://doi.org/10.1186/s42649-021-00066-7Clay mineralProtein bindingHomology modelingMutationAFMAdhesion force |
| spellingShingle | Renukaradhya K. Math Nagakumar Bharatham Palaksha K. Javaregowda Han Dae Yun Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces Applied Microscopy Clay mineral Protein binding Homology modeling Mutation AFM Adhesion force |
| title | Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces |
| title_full | Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces |
| title_fullStr | Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces |
| title_full_unstemmed | Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces |
| title_short | Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces |
| title_sort | role of cel5h protein surface amino acids in binding with clay minerals and measurements of its forces |
| topic | Clay mineral Protein binding Homology modeling Mutation AFM Adhesion force |
| url | https://doi.org/10.1186/s42649-021-00066-7 |
| work_keys_str_mv | AT renukaradhyakmath roleofcel5hproteinsurfaceaminoacidsinbindingwithclaymineralsandmeasurementsofitsforces AT nagakumarbharatham roleofcel5hproteinsurfaceaminoacidsinbindingwithclaymineralsandmeasurementsofitsforces AT palakshakjavaregowda roleofcel5hproteinsurfaceaminoacidsinbindingwithclaymineralsandmeasurementsofitsforces AT handaeyun roleofcel5hproteinsurfaceaminoacidsinbindingwithclaymineralsandmeasurementsofitsforces |