Breast cancer is one of the most common cancers in women around the globe Tamoxifen is used for the last 40 years as an endocrine therapy for breast cancer. This resulted in the reduction of mortality rate by 30% and it still remains one of the most effective therapies against breast cancer. However, resistance against tamoxifen is still one of the major hurdles in the effective management of breast cancer. Intense research has been conducted in the past decade to further explore its resistance mechanism, but still a lot of research will be needed to effectively alleviate this problem. Several biochemical factors and molecular pathways, such as the modulation of ER signaling, upregulation of growth factors had been observed as key factors for tamoxifen resistance (TR). After, initial therapy of five to ten years, breast cancer patients develops resistance towards this drug. The resistance leads to the development of other cancers like uterine cancer. Here, we briefly explore all the molecular events related to tamoxifen resistance and focus on its mechanism of action as well as other pharmacological approaches to better its beneficial effects in the treatment of breast carcinoma.

Troponin T (TnT), a thin filament myofibrillar protein, is essential for the Ca 2؉ regulation of striated muscle contraction in vertebrates, both in vivo and in vitro. To understand the role of TnT in this process, its interaction with two other troponin components, troponin I (TnI) and troponin C (TnC) was examined by using the yeast two hybrid system, which is a genetic approach to detect protein-protein interactions. Computer assisted analysis of phylogenetically distant TnT amino acid sequences unveiled a highly conserved protein domain that is characterized by a heptad repeat (HR) motif with a potential for ␣-helical coiled coil formation. A similar, potentially coiled coil forming domain is also conserved in all known TnI sequences. These protein motifs appeared to be the regions where TnI-TnT interaction may take place. Deletions and point mutations in TnT, which disrupted its HR motif, severely reduced or abolished TnI binding, but binding to TnC was not affected, indicating that the TnT-TnI and TnT-TnC binary interactions can be uncoupled. Remarkably, the truncated fragments of TnT and TnI in which the HR motifs were retained showed binary interaction in the yeast two hybrid system. It was also observed that the formation of the TnT-TnI heterodimers is favored over the homodimers TnT-TnT and TnI-TnI. These results indicate that the evolutionarily conserved HR motifs may play a role in TnT-TnI dimerization, presumably through the formation of ␣-helical coiled coils.

The interaction between troponin I (TnI) and troponin T (TnT) remains the least understood binary interaction among the regulatory proteins of vertebrate striated muscle. To identify the specific binding domains of TnI and TnT and to evaluate the interactions of TnT with troponin C and tropomyosin (Tm), we generated an NH2-terminal fragment of human fast skeletal beta TnT (TnT1-201; residues 1-201) using site-directed mutagenesis. The mutant protein failed to bind to rabbit skeletal muscle TnI as judged by HPLC, showed reduced TnC binding and reduced ternary troponin (Tn) complex formation, and exhibited a much reduced Ca2+ sensitivity in the reconstituted regulatory system. It is shown that the amount of Tn complex formed by TnT1-201 rather than the activity of the mutant Tn complex affected this Ca2+ sensitivity. Binding of the mutant to Tm was similar to that of intact TnT. These results support the view that the COOH-terminal segment of TnT is necessary for binding to TnI and TnC and Ca2+ sensitivity in the thin filament, whereas its NH2-terminus strongly binds to Tm. To identify the regions of TnI which bind to muscle TnT, we used four recombinant fragments of fast skeletal muscle TnI containing amino acid residues 1-94 (TnI1-94), 1-120 (TnI1-120), 96-181 (TnI96-181), and 122-181 (TnI122-181) and a synthetic peptide, TnI98-114, containing residues 98-114 corresponding to the inhibitory region. Only TnI1-120 showed weak binding to TnT but not to TnT1-201. These results suggest that (i) a region within the NH2-terminal 120 residues of TnI interacts with TnT and (ii) the COOH-terminal residues 202-258 of TnT contain the interaction site of TnI. Overall, our results also imply that residues 159-201 constitute the smallest region of TnT which contributes to the Ca2+ sensitivity of actoS1 ATPase in a reconstituted regulatory system.

Contraction of vertebrate striated muscle is regulated by the strong Ca(2+)-dependent interaction between troponin I (TnI) and troponin C (TnC). To critically evaluate this interaction, we generated four recombinant deletion fragments of rabbit fast skeletal TnI: the NH2-terminal fragment (TnI1-94), the NH2 terminus and the inhibitory region (TnI1-120), the inhibitory region and the COOH terminus (TnI96-181), and the COOH-terminal fragment (TnI122-181) containing amino acid residues 1-94, 1-120, 96-181, and 122-181, respectively. Native TnC and seven thiol mutants, containing single cysteine residues in the two globular domains and in the central helix of TnC, e.g., Cys-12, Cys-21, Cys-57, Cys-89, Cys-122, Cys-133, and Cys-158, were labeled with 4-maleimidobenzophenone, and their interaction with the recombinant TnI fragments and the synthetic inhibitory peptide (TnI98-114, residues 98-114) was studied by photo-cross-linking. Extensive cross-linking occurred between various domains of TnC and TnI. The cross-linking patterns (a) showed that both NH2- and COOH-terminal fragments of TnI are accessible to both of the globular domains of TnC, (b) indicated that linkage of the NH2- and COOH-terminal sequences to the inhibitory region of TnI (TnIir) caused marked enhancement of cross-linking with native TnC and all seven thiol mutants, and (c) identified the region in TnC where TnIir binds as that containing residues 98, 133, 158, and 57. Thus, the results suggest that TnI and TnC may adopt flexible and dynamic conformations in which multiple interactions involving various domains of the two polypeptides occur and TnIir acting as a linker facilitates these interactions. The interaction of TnI and its fragments with actin, TnC, and TnT, considered together with the biological activity indicates that residues 96-120 represent a key structural and functional region of TnI. Whereas the NH2-terminal region of TnI stabilizes binding to TnC and TnT, the COOH-terminal region stabilizes TnC and actin binding.