1. Click "Predict" tab to submit a job in MPTherm-pred server.
2. Enter the protein data bank (PDB) accession number.
3. Specify the chain ID for the respective PDB code.
4. Enter missense mutation in the following format: Wildtype residue-space-Position in the protein sequence-space-Mutant residue.

Example: A227Y. Where, A and Y is a wild-type and mutant residue of mutation, respectively. 227 denotes the position of wild-type residue in the sequence.

5. Specify the function of a protein. Based on functions, mutations in the membrane-spanning region were classified into three classes such as Transporters & others, receptors and Unknown (combined both the classes).

If the user is not specifying the functional class, MPTherm-pred will consider that given mutation as the unknown functional class. For more accurate results, we suggest that users should specify the functional information. The functional information of a protein is available in UniProt database.