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Characterization of a core fragment of the rhesus monkey TRIM5α protein.

Kar AK, Mao Y, Bird G, Walensky L, Sodroski J - BMC Biochem. (2011)

Bottom Line: This BCCL2 protein formed dimers and higher-order oligomers in solution.Approximately 40% of the BCCL2 secondary structure consisted of alpha helices.These results indicate that the B-box 2, coiled-coil and linker 2 regions of TRIM5α form a core dimerization motif that exhibits a high level of alpha-helical content.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA.

ABSTRACT

Background: Like all tripartite motif (TRIM) proteins, the retroviral restriction factor TRIM5α consists of RING, B-box 2 and coiled-coil domains, with a C-terminal B30.2(SPRY) domain. Although structures have been determined for some individual TRIM domains, the structure of an intact TRIM protein is unknown.

Results: Here, we express and characterize a protease-resistant 29-kD core fragment containing the B-box 2, coiled coil and adjacent linker (L2) region of TRIM5α. This BCCL2 protein formed dimers and higher-order oligomers in solution. Approximately 40% of the BCCL2 secondary structure consisted of alpha helices. Partial loss of alpha-helical content and dissociation of dimers occurred at 42°C, with the residual alpha helices remaining stable up to 80°C.

Conclusions: These results indicate that the B-box 2, coiled-coil and linker 2 regions of TRIM5α form a core dimerization motif that exhibits a high level of alpha-helical content.

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Related in: MedlinePlus

Dynamic light scattering analysis of the LLER protein. The polydispersity (%Pd) and size (mean and mode, in nm) of the purified LLER protein were estimated by dynamic light scattering. A 12-μl sample of a 500 μg/ml solution of the LLER protein in peak 1 (A) and peak 2 (B) were analyzed by a temperature-controlled Zetasizer Nano-S dynamic light-scattering instrument at 20°C.
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Figure 6: Dynamic light scattering analysis of the LLER protein. The polydispersity (%Pd) and size (mean and mode, in nm) of the purified LLER protein were estimated by dynamic light scattering. A 12-μl sample of a 500 μg/ml solution of the LLER protein in peak 1 (A) and peak 2 (B) were analyzed by a temperature-controlled Zetasizer Nano-S dynamic light-scattering instrument at 20°C.

Mentions: We conducted additional dynamic light-scattering analysis of the two purified LLER protein fractions from size-exclusion chromatography (Figure 6). The protein in Peak 1 exhibited a diameter of 11.6 nm and a polydispersity of 22%, and the protein in Peak 2 exhibited a diameter of 8.6 nm and a polydispersity of 45%.


Characterization of a core fragment of the rhesus monkey TRIM5α protein.

Kar AK, Mao Y, Bird G, Walensky L, Sodroski J - BMC Biochem. (2011)

Dynamic light scattering analysis of the LLER protein. The polydispersity (%Pd) and size (mean and mode, in nm) of the purified LLER protein were estimated by dynamic light scattering. A 12-μl sample of a 500 μg/ml solution of the LLER protein in peak 1 (A) and peak 2 (B) were analyzed by a temperature-controlled Zetasizer Nano-S dynamic light-scattering instrument at 20°C.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3025952&req=5

Figure 6: Dynamic light scattering analysis of the LLER protein. The polydispersity (%Pd) and size (mean and mode, in nm) of the purified LLER protein were estimated by dynamic light scattering. A 12-μl sample of a 500 μg/ml solution of the LLER protein in peak 1 (A) and peak 2 (B) were analyzed by a temperature-controlled Zetasizer Nano-S dynamic light-scattering instrument at 20°C.
Mentions: We conducted additional dynamic light-scattering analysis of the two purified LLER protein fractions from size-exclusion chromatography (Figure 6). The protein in Peak 1 exhibited a diameter of 11.6 nm and a polydispersity of 22%, and the protein in Peak 2 exhibited a diameter of 8.6 nm and a polydispersity of 45%.

Bottom Line: This BCCL2 protein formed dimers and higher-order oligomers in solution.Approximately 40% of the BCCL2 secondary structure consisted of alpha helices.These results indicate that the B-box 2, coiled-coil and linker 2 regions of TRIM5α form a core dimerization motif that exhibits a high level of alpha-helical content.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02115, USA.

ABSTRACT

Background: Like all tripartite motif (TRIM) proteins, the retroviral restriction factor TRIM5α consists of RING, B-box 2 and coiled-coil domains, with a C-terminal B30.2(SPRY) domain. Although structures have been determined for some individual TRIM domains, the structure of an intact TRIM protein is unknown.

Results: Here, we express and characterize a protease-resistant 29-kD core fragment containing the B-box 2, coiled coil and adjacent linker (L2) region of TRIM5α. This BCCL2 protein formed dimers and higher-order oligomers in solution. Approximately 40% of the BCCL2 secondary structure consisted of alpha helices. Partial loss of alpha-helical content and dissociation of dimers occurred at 42°C, with the residual alpha helices remaining stable up to 80°C.

Conclusions: These results indicate that the B-box 2, coiled-coil and linker 2 regions of TRIM5α form a core dimerization motif that exhibits a high level of alpha-helical content.

Show MeSH
Related in: MedlinePlus