Purification of tubulin and microtubules from mouse brain

Purified tubulin or microtubules are widely used for in vitro assays related to their function, such as activities of their modifying enzymes and motor protein activity. This paper gave a review for purification tubulin and microtubules (MTs) using two different methods. One method of purification MTs depended on polymerization and depolymerization. MTs were polymerized at 37 °C with taxol in the presence. Another method is to purify tubulin by affinity column chromatography. Stu2p, a kind of MAPs, is able to bind with C-terminal of tubulin. Recombinant Stu2p was purified from expressing bacteria. Then Stu2p was coupled with NHS-activated resins, which were used to purify tubulin from mouse brain. Tubulins purified in this way can also be polymerized at 37 °C.

Proteomic identification of galectin-11 and 14 ligands fromHaemonchus contortus

Haemonchus contortusis the most pathogenic nematode of small ruminants. Infection in sheep and goats results in anaemia that decreases animal productivity and can ultimately cause death. The involvement of ruminant-specific galectin-11 (LGALS-11) and galectin-14 (LGALS-14) has been postulated to play important roles in protective immune responses against parasitic infection; however, their ligands are unknown. In the current study, LGALS-11 and LGALS-14 ligands inH. contortuswere identified from larval (L4) and adult parasitic stages extracts using immobilised LGALS-11 and LGALS-14 affinity column chromatography and mass spectrometry. Both LGALS-11 and LGALS-14 bound more putative protein targets in the adult stage ofH. contortus(43 proteins) when compared to the larval stage (two proteins). Of the 43 proteins identified in the adult stage, 34 and 35 proteins were bound by LGALS-11 and LGALS-14, respectively, with 26 proteins binding to both galectins. Interestingly, hematophagous stage-specific sperm-coating protein and zinc metalloprotease (M13), which are known vaccine candidates, were identified as putative ligands of both LGALS-11 and LGALS-14. The identification of glycoproteins ofH. contortusby LGALS-11 and LGALS-14 provide new insights into host-parasite interactions and the potential for developing new interventions.

Proteomic identification of Galectin-11 and 14 ligands from Haemonchus contortus

Haemonchus contortus is the most pathogenic nematode of small ruminants. Infection in sheep and goats results in anaemia that decreases animal productivity and can ultimately cause death. The involvement of ruminant-specific galectin-11 (LGALS-11) and galectin-14 (LGALS-14) has been postulated to play important roles in protective immune responses against parasitic infection; however, their ligands are unknown. In the current study, LGALS-11 and LGALS-14 ligands in H. contortus were identified from larval (L4) and adult parasitic stages extracts using immobilised LGALS-11 and LGALS-14 affinity column chromatography and mass spectrometry. Both LGALS-11 and LGALS-14 bound more putative protein targets in the adult stage of H. contortus (43 proteins) when compared to the larval stage (2 proteins). Of the 43 proteins identified in the adult stage, 34 and 35 proteins were bound by LGALS-11 and LGALS-14, respectively, with 26 proteins binding to both galectins. Interestingly, hematophagous stage-specific sperm-coating protein and zinc metalloprotease (M13), which are known vaccine candidates, were identified as putative ligands of both LGALS-11 and LGALS-14. The identification of glycoproteins of H. contortus by LGALS-11 and LGALS-14 provide new insights into host-parasite interactions and the potential for developing new interventions.

Proteomic identification of Galectin-11 and 14 ligands from Haemonchus contortus

Haemonchus contortus is the most pathogenic nematode of small ruminants. Infection in sheep and goats results in anaemia that decreases animal productivity and can ultimately cause death. The involvement of ruminant-specific galectin-11 (LGALS-11) and galectin-14 (LGALS-14) has been postulated to play important roles in protective immune responses against parasitic infection; however, their ligands are unknown. In the current study, LGALS-11 and LGALS-14 ligands in H. contortus were identified from larval (L4) and adult parasitic stages extracts using immobilised LGALS-11 and LGALS-14 affinity column chromatography and mass spectrometry. Both LGALS-11 and LGALS-14 bound more putative protein targets in the adult stage of H. contortus (43 proteins) when compared to the larval stage (2 proteins). Of the 43 proteins identified in the adult stage, 34 and 35 proteins were bound by LGALS-11 and LGALS-14, respectively, with 26 proteins binding to both galectins. Interestingly, hematophagous stage-specific sperm-coating protein and zinc metalloprotease (M13), which are known vaccine candidates, were identified as putative ligands of both LGALS-11 and LGALS-14. The identification of glycoproteins of H. contortus by LGALS-11 and LGALS-14 provide new insights into host-parasite interactions and the potential for developing new interventions.

CLONING AND EXPRESSION OF VACCINIA VIRUS PROTEIN (B18) IN HETEROLOGOUS HOST

B18 is an immunoglobulin superfamily glycoprotein produced by vaccinia virus which resembles to interferon alpha receptors. It acts both in solution and also when associated with the cell surface. This B18 protein was expressed in heterologous host and purified based on affinity column chromatography. The purified B18 protein has been evaluated for biological activity in Pig Kidney- 15 cells (PK-15) which has been reported to have α –interferon receptors and signaling mechanism. The CPE was more than 90% in the presence of protein and around 50% in the absence. B-18 can enhance the infectivity of FMDV to cells having α –interferon receptors. This provides the basis for developing new strategies for improving attenuated vaccines and its capacity to act as an enhancer protein.