Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, -5

Matched in the w>database search indicated with asterisks. Panels correspond to a spot from the 20S() and 20S() proteasomes, respectively.<p><b>Copyright information:</b></p><p>Taken from "Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, "</p><p>http://www.biomedcentral.com/1471-2091/9/20</p><p>BMC Biochemistry 2008;9():20-20.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2515314.</p><p></p

Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, -10

Ome, 1 mM EDTA-Tris, pH 8.5, 10% DMSO, and 10 μM peptidyl substrates (: Suc-LLVY-MCA for chymotrypsin-like activity; : Z-LLE-MCA for PGPH activity; : Boc-LRR-MCA for trypsin-like activity) and increasing concentrations of linolenic acid at 37°C for 1 h. The reactions were started, stopped as described in methods. Symbols represent the wild-type strain (○) and the mutant (●). Values are means ± SD of three independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, "</p><p>http://www.biomedcentral.com/1471-2091/9/20</p><p>BMC Biochemistry 2008;9():20-20.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2515314.</p><p></p

Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, -9

50 mM Tris-HCl, pH 7.6, 1 μg/ml 20S proteasome, and 10 μM peptidyl substrates (: Suc-LLVY-MCA for chymotrypsin-like activity; : Z-LLE-MCA for PGPH activity; : Boc-LRR-MCA for trypsin-like activity) and increasing concentrations of SDS at 37°C for 1 h. The reactions were started and stopped as described in methods. Symbols represent the wild-type strain (○) and the mutant (●). Values are means ± SD of three independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, "</p><p>http://www.biomedcentral.com/1471-2091/9/20</p><p>BMC Biochemistry 2008;9():20-20.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2515314.</p><p></p

Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, -0

50 mM Tris-HCl, pH 7.6, 1 μg/ml 20S proteasome, and 10 μM peptidyl substrates (: Suc-LLVY-MCA for chymotrypsin-like activity; : Z-LLE-MCA for PGPH activity; : Boc-LRR-MCA for trypsin-like activity) and increasing concentrations of SDS at 37°C for 1 h. The reactions were started and stopped as described in methods. Symbols represent the wild-type strain (○) and the mutant (●). Values are means ± SD of three independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, "</p><p>http://www.biomedcentral.com/1471-2091/9/20</p><p>BMC Biochemistry 2008;9():20-20.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2515314.</p><p></p

Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, -1

Some, 1 mM EDTA-Tris, pH 8.5, 10% DMSO, and 10 μM peptidyl substrates (: Suc-LLVY-MCA for chymotrypsin-like activity; : Z-LLE-MCA for PGPH activity; : Boc-LRR-MCA for trypsin-like activity) and increasing concentrations of linolenic acid at 37°C for 1 h. The reactions were started, stopped as described in methods. Symbols represent the wild-type strain (○) and the mutant (●). Values are means ± SD of three independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, "</p><p>http://www.biomedcentral.com/1471-2091/9/20</p><p>BMC Biochemistry 2008;9():20-20.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2515314.</p><p></p

Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, -2

20S proteasome, and 10 μM of peptidyl substrates (: Suc-LLVY-MCA for chymotrypsin-like activity; : Z-LLE-MCA for PGPH activity; : Boc-LRR-MCA for trypsin-like activity) at 37°C for 1 h. Reactions were started and stopped as described in methods. Symbols represent the wild-type strain (○) and the mutant (●). Values are means ± SD for three independent experiments.<p><b>Copyright information:</b></p><p>Taken from "Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, "</p><p>http://www.biomedcentral.com/1471-2091/9/20</p><p>BMC Biochemistry 2008;9():20-20.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2515314.</p><p></p

Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, -7

Erred to a PVDF membrane. The α1- and α7-subunits were detected using their respective antibodies as described in the methods. () Cells from wild-type and the mutant (wt, ) grown at 25°C (OD= 1.0) were harvested and the cell extracts were applied to a Q-sepharose column. Active Q-sepharose fractions were concentrated and treated with (AP+) or without (AP-) alkaline phosphatase as described in the methods. The samples were then subjected to SDS-PAGE in a 12% polyacrylamide gel. The α7-subunit was detected by immunostaining with anti-α7 antibody [,].<p><b>Copyright information:</b></p><p>Taken from "Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, "</p><p>http://www.biomedcentral.com/1471-2091/9/20</p><p>BMC Biochemistry 2008;9():20-20.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2515314.</p><p></p

Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, -8

Ltures were then immediately shifted to 37°C and incubated for several hours as indicated. Cell extracts were applied to a Q-sepharose column. The molecular weight level of the α7-subunits in both strains produced during the indicated incubation period was assayed by Western blotting using anti-α7 antibody.<p><b>Copyright information:</b></p><p>Taken from "Functional and biochemical characterization of the 20S proteasome in a yeast temperature-sensitive mutant, "</p><p>http://www.biomedcentral.com/1471-2091/9/20</p><p>BMC Biochemistry 2008;9():20-20.</p><p>Published online 21 Jul 2008</p><p>PMCID:PMC2515314.</p><p></p