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Protein-protein interactions govern almost all biological processes and the underlying functions of proteins. The interaction sites of protein depend on the 3D structure which in turn depends on the amino acid sequence. Hence, prediction of protein function from its primary sequence is an important and challenging task in bioinformatics. Identification of the amino acids (hot spots) that leads to the characteristic frequency signifying a particular biological function is really a tedious job in proteomic signal processing. In this paper, we have proposed a new promising technique for identification of hot spots in proteins using an efficient time-frequency filtering approach known as the S-transform filtering. The S-transform is a powerful linear time-frequency representation and is especially useful for the filtering in the time-frequency domain. The potential of the new technique is analyzed in identifying hot spots in proteins and the result obtained is compared with the existing methods. The results demonstrate that the proposed method is superior to its counterparts and is consistent with results based on biological methods for identification of the hot spots. The proposed method also reveals some new hot spots which need further investigation and validation by the biological community.