Role of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces

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...

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Autores principales: Renukaradhya K. Math, Nagakumar Bharatham, Palaksha K. Javaregowda, Han Dae Yun
Formato: article
Lenguaje:EN
Publicado: SpringerOpen 2021
Materias:
Clay mineral
Protein binding
Homology modeling
Mutation
AFM
Adhesion force
Microscopy
QH201-278.5
Acceso en línea:https://doaj.org/article/a99cb17f71c14186865bf020012d35cc
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spelling oai:doaj.org-article:a99cb17f71c14186865bf020012d35cc2021-11-14T12:33:05ZRole of Cel5H protein surface amino acids in binding with clay minerals and measurements of its forces10.1186/s42649-021-00066-72287-4445https://doaj.org/article/a99cb17f71c14186865bf020012d35cc2021-11-01T00:00:00Zhttps://doi.org/10.1186/s42649-021-00066-7https://doaj.org/toc/2287-4445Abstract 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.Renukaradhya K. MathNagakumar BharathamPalaksha K. JavaregowdaHan Dae YunSpringerOpenarticleClay mineralProtein bindingHomology modelingMutationAFMAdhesion forceMicroscopyQH201-278.5ENApplied Microscopy, Vol 51, Iss 1, Pp 1-10 (2021)
institution DOAJ
collection DOAJ
language EN
topic Clay mineral
Protein binding
Homology modeling
Mutation
AFM
Adhesion force
Microscopy
QH201-278.5
spellingShingle Clay mineral
Protein binding
Homology modeling
Mutation
AFM
Adhesion force
Microscopy
QH201-278.5
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
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.
format article
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
title 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_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_sort role of cel5h protein surface amino acids in binding with clay minerals and measurements of its forces
publisher SpringerOpen
publishDate 2021
url https://doaj.org/article/a99cb17f71c14186865bf020012d35cc
work_keys_str_mv AT renukaradhyakmath roleofcel5hproteinsurfaceaminoacidsinbindingwithclaymineralsandmeasurementsofitsforces
AT nagakumarbharatham roleofcel5hproteinsurfaceaminoacidsinbindingwithclaymineralsandmeasurementsofitsforces
AT palakshakjavaregowda roleofcel5hproteinsurfaceaminoacidsinbindingwithclaymineralsandmeasurementsofitsforces
AT handaeyun roleofcel5hproteinsurfaceaminoacidsinbindingwithclaymineralsandmeasurementsofitsforces
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