caa a22 4500 605512191 CHVBK 20210128100654.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00894-015-2807-x doi (NATIONALLICENCE)springer-10.1007/s00894-015-2807-x Understanding the roles of Lys33 and Arg45 in the binding-site stability of LjLTP10, an LTP related to drought stress in Lotus japonicus [Elektronische Daten] [Felipe Valenzuela-Riffo, Gerardo Tapia, Carolina Parra-Palma, Luis Morales-Quintana] In Lotus japonicus, as in most plants, long-chain fatty acids are important components of cuticular wax, one of the principal functions of which is to act as a barrier to water loss in response to drought stress. It is thought that lipid transfer proteins (LTPs) are involved in the process of cuticle formation. We previously described LjLTP10 as an LTP involved in cuticle formation during acclimation response to drought stress in L. japonicus. The structural model of LjLTP10 had two residues (K33 and R45) in the hydrophobic cavity, although the role of these residues was unclear. In the present work, we investigated the molecular mechanism involved in the transport of lipid precursors in L. japonicus and clarified the importance of the residues K33 and R45. First, in silico site-directed mutagenesis studies were carried out on the LjLTP10 structure. Structural analysis showed that LjLTP10 mutants possess similar structures but their hydrophobic cavities are somewhat different. Unfavorable energies for the interactions of the mutant proteins with different ligands were found by molecular docking and molecular dynamics simulations. We also examined the contributions of energetic parameters to the free energy of the protein-ligand complex using the MM-GBSA method. Results showed that the different complexes present similar, favorable van der Waals interactions, whereas electrostatic interactions were not favored in the mutant structures. Our study indicates that the residues K33 and R45 play a crucial role in maintaining the binding pocket structure required for lipid transport. Springer-Verlag Berlin Heidelberg, 2015 Lipid transfer protein nationallicence In silico site-directed mutagenesis nationallicence Molecular dynamics simulations nationallicence MM-GBSA nationallicence Lotus japonicus nationallicence Valenzuela-Riffo Felipe Escuela de Ingeniería en Bioinformática, Facultad de Ingeniería, Universidad de Talca, Casilla 747, Talca, Chile aut Tapia Gerardo Unidad de Recursos Genéticos, Instituto de Investigaciones Agropecuarias, INIA-Quilamapu, Chillán, Chile aut Parra-Palma Carolina Laboratorio de Fisiología Vegetal y Genética Molecular, Instituto de Ciencias Biológicas, Universidad de Talca, Casilla 747, Talca, Chile aut Morales-Quintana Luis Laboratorio de Fisiología Vegetal y Genética Molecular, Instituto de Ciencias Biológicas, Universidad de Talca, Casilla 747, Talca, Chile aut Journal of Molecular Modeling Springer Berlin Heidelberg 21/10(2015-10-01), 1-11 1610-2940 21:10<1 2015 21 894 https://doi.org/10.1007/s00894-015-2807-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00894-015-2807-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Valenzuela-Riffo Felipe Escuela de Ingeniería en Bioinformática, Facultad de Ingeniería, Universidad de Talca, Casilla 747, Talca, Chile aut NATIONALLICENCE 700 1- Tapia Gerardo Unidad de Recursos Genéticos, Instituto de Investigaciones Agropecuarias, INIA-Quilamapu, Chillán, Chile aut NATIONALLICENCE 700 1- Parra-Palma Carolina Laboratorio de Fisiología Vegetal y Genética Molecular, Instituto de Ciencias Biológicas, Universidad de Talca, Casilla 747, Talca, Chile aut NATIONALLICENCE 700 1- Morales-Quintana Luis Laboratorio de Fisiología Vegetal y Genética Molecular, Instituto de Ciencias Biológicas, Universidad de Talca, Casilla 747, Talca, Chile aut NATIONALLICENCE 773 0- Journal of Molecular Modeling Springer Berlin Heidelberg 21/10(2015-10-01), 1-11 1610-2940 21:10<1 2015 21 894