TSpred: a web server for the rational design of temperature-sensitive mutants.
Temperature sensitive (Ts) mutants of proteins provide experimentalists with a powerful and reversible way of conditionally expressing genes. The technique has been widely used in determining the role of gene and gene products in several cellular processes. Traditionally, Ts mutants are generated by random mutagenesis and then selected though laborious large-scale screening. Our web server, TSpred (http://mspc.bii.a-star.edu.sg/TSpred/), now enables users to rationally design Ts mutants for their proteins of interest. TSpred uses hydrophobicity and hydrophobic moment, deduced from primary sequence and residue depth, inferred from 3D structures to predict/identify buried hydrophobic residues. Mutating these residues leads to the creation of Ts mutants. Our method has been experimentally validated in 36 positions in six different proteins. It is an attractive proposition for Ts mutant engineering as it proposes a small number of mutations and with high precision. The accompanying web server is simple and intuitive to use and can handle proteins and protein complexes of different sizes.
Bioinformatics Institute, 30 Biopolis Street, #07-01, Matrix, Singapore 138671 School of Computer Engineering, Nanyang Technological University, Singapore 639798.
Kuan Pern
KP
Tan
Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560012, India.
Shruti
S
Khare
Molecular Biophysics Unit, Indian Institute of Science, Bangalore 560012, India madhusudhan@iiserpune.ac.in.
Raghavan
R
Varadarajan
Bioinformatics Institute, 30 Biopolis Street, #07-01, Matrix, Singapore 138671 Indian Institute of Science Education and Research, Pune 411008, India Department of Biological Sciences, National University of Singapore, Singapore 117543 madhusudhan@iiserpune.ac.in.
Mallur Srivatsan
MS
Madhusudhan
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D020407:Internet; D009113:Muramidase / Q000737:chemistry / Q000235:genetics; D009154:Mutation; D011506:Proteins / Q000737:chemistry / Q000235:genetics*; D012984:Software; D013696:Temperature
D001426:Bacterial Proteins; C068356:CcdB protein, Plasmid F; D011506:Proteins; D009113:Muramidase