Journal: Cardiovascular Research

Article Title: Ecstasy produces left ventricular dysfunction and oxidative stress in rats

doi: 10.1093/cvr/cvn129

Figure Lengend Snippet: 3,4-methylenedioxymethamphetamine increases tyrosine nitration of SERCA-2a. (A) Nitrotyrosine immunoprecipitates from saline (n = 5 pooled) and 3,4-methylenedioxymethamphetamine (n = 6 pooled) left ventricular lysates were probed by western blot analysis using SERCA-2a antibody. (B) SERCA-2a protein levels normalized to GAPDH (n = 5). (C) SERCA-2a protein levels normalized to phospholamban (n = 5).

Article Snippet: Immunoprecipitated proteins were subjected to western blot analysis with SERCA-2a (Biomol) antibodies.

Techniques: Nitration, Western Blot

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Structural basis for the high Ca 2+ affinity of the ubiquitous SERCA2b Ca 2+ pump

doi: 10.1073/pnas.0906797106

Figure Lengend Snippet: Functional effects of the luminal residues of the 2b-tail. (A) Dose–response curve of luminal peptide LE (n = 3). (B) Normalized Ca2+-dependent ATPase activity of SERCA2a in the presence (○) and absence (●) of 1.2 mM LE (n = 3). (C) Average K0.5 values for Ca2+ of the alanine mutants of the luminal 2b-tail (n = 3–5). P < 0.05 vs. SERCA2b (*) or SERCA2a (#). (D) Partial proteinase K digestion of SERCA isoforms in the presence or absence of 50 μM CaCl2, 1 mM EGTA, or 4 mM LE peptide. Arrow marks E1 stabilization of SERCA2b in the absence of Ca2+. S1a, SERCA1a; S2a, SERCA2a; S2b, SERCA2b.

Article Snippet: Note that this site is only properly formed in E1–2Ca and E1∼P, is slightly transformed in E2-P, and is completely lacking in E2 ( C and S3 d ), in agreement with the expected stabilization of E1 by LE. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. 4. caption a7 Residues MFWS occupy a luminal docking site formed by L1–2, L3–4, and L7–8. ( A ) Docking of the MFWS tetrapeptide to the luminal region of SERCA2a (homology model in E1–2Ca) using Autodock (−7.85 kcal/mol).

Techniques: Functional Assay, Activity Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Structural basis for the high Ca 2+ affinity of the ubiquitous SERCA2b Ca 2+ pump

doi: 10.1073/pnas.0906797106

Figure Lengend Snippet: Functional effect of TM11 in SERCA2b and SERCA1a2b chimera. Average K0.5 values of various SERCA2b mutants (A) and SERCA1a2b chimera (B) (n = 7–10). P < 0.05 vs. SERCA2b (*), SERCA2a (#), or SERCA1a ($).

Article Snippet: Note that this site is only properly formed in E1–2Ca and E1∼P, is slightly transformed in E2-P, and is completely lacking in E2 ( C and S3 d ), in agreement with the expected stabilization of E1 by LE. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. 4. caption a7 Residues MFWS occupy a luminal docking site formed by L1–2, L3–4, and L7–8. ( A ) Docking of the MFWS tetrapeptide to the luminal region of SERCA2a (homology model in E1–2Ca) using Autodock (−7.85 kcal/mol).

Techniques: Functional Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Structural basis for the high Ca 2+ affinity of the ubiquitous SERCA2b Ca 2+ pump

doi: 10.1073/pnas.0906797106

Figure Lengend Snippet: The 2b-tail interacts with loop L7–8. (A) Average K0.5 values of SERCA1a2b chimera with replacement of luminal loops L5–6, L7–8, and/or L9–10 (n = 4–6). (B) Average K0.5 values of L7–8 SERCA2b mutants (n = 6–10). N terminal, middle, and C terminal indicate the position of the multiple mutations on L7–8. P < 0.05 vs. SERCA2b (*), SERCA2a (#), SERCA1a ($), or SERCA1a2b (°).

Article Snippet: Note that this site is only properly formed in E1–2Ca and E1∼P, is slightly transformed in E2-P, and is completely lacking in E2 ( C and S3 d ), in agreement with the expected stabilization of E1 by LE. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. 4. caption a7 Residues MFWS occupy a luminal docking site formed by L1–2, L3–4, and L7–8. ( A ) Docking of the MFWS tetrapeptide to the luminal region of SERCA2a (homology model in E1–2Ca) using Autodock (−7.85 kcal/mol).

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Structural basis for the high Ca 2+ affinity of the ubiquitous SERCA2b Ca 2+ pump

doi: 10.1073/pnas.0906797106

Figure Lengend Snippet: Residues MFWS occupy a luminal docking site formed by L1–2, L3–4, and L7–8. (A) Docking of the MFWS tetrapeptide to the luminal region of SERCA2a (homology model in E1–2Ca) using Autodock (−7.85 kcal/mol). MFWS is indicated in gray spheres. All residues within a radius of 4 Å of MFWS are shown as sticks. Residues T86, V283, H284, and Y294 are labeled. (B) Average K0.5 values of SERCA2b (white) and SERCA2a (dark gray) WT and luminal mutants (speckled) (n = 3–6). P < 0.05 vs. SERCA2b (*) or SERCA2a (#). (C) Luminal docking site in the two main conformations of SERCA2a (homology models based on SERCA1a structures (3) 1SU4 (E1) and 2AGV (E2). T86, V283, H284, Y294, and K876 are shown as black sticks.

Article Snippet: Note that this site is only properly formed in E1–2Ca and E1∼P, is slightly transformed in E2-P, and is completely lacking in E2 ( C and S3 d ), in agreement with the expected stabilization of E1 by LE. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. 4. caption a7 Residues MFWS occupy a luminal docking site formed by L1–2, L3–4, and L7–8. ( A ) Docking of the MFWS tetrapeptide to the luminal region of SERCA2a (homology model in E1–2Ca) using Autodock (−7.85 kcal/mol).

Techniques: Labeling

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Structural basis for the high Ca 2+ affinity of the ubiquitous SERCA2b Ca 2+ pump

doi: 10.1073/pnas.0906797106

Figure Lengend Snippet: TM11 interaction site. (A) Average K0.5 values of TM10 mutants (n = 4–9). P < 0.05 vs. SERCA2b (*) or SERCA2a (#). (B) NMR structure of TM11 in DMSO/CDCl3. Red bar shows N-terminal cap between the side chain of S1015 and the peptide backbone of F1018. (C) Detailed surface view of the docking model of TM11 (yellow) in the region TM7, TM8, TM9, TM10 (green). Residues from the IA are shown in light oragne (SERCA) and dark orange (TM11). (D) Surface view of the IA (orange) of TM11 on TM7, TM8, TM9, TM10 (green). All indicated residues of the IA are conserved in SERCA2. Residues indicated by red labels are not identical but conserved in SERCA1a, and those indicated in blue are not conserved in SERCA1a. (E) Surface view of the IA (orange) on TM11 (yellow). Residues identical in the 2b-tail of vertebrates are labeled in black; not identical, but conserved are in red.

Article Snippet: Note that this site is only properly formed in E1–2Ca and E1∼P, is slightly transformed in E2-P, and is completely lacking in E2 ( C and S3 d ), in agreement with the expected stabilization of E1 by LE. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Fig. 4. caption a7 Residues MFWS occupy a luminal docking site formed by L1–2, L3–4, and L7–8. ( A ) Docking of the MFWS tetrapeptide to the luminal region of SERCA2a (homology model in E1–2Ca) using Autodock (−7.85 kcal/mol).

Techniques: Labeling

Journal: Cardiovascular Diagnosis and Therapy

Article Title: In vivo effects of nitrosyl hydrogen on cardiac function and sarcoplasmic reticulum calcium pump (SERCA2a) in rats with heart failure after myocardial infarction

doi: 10.21037/cdt-20-201

Figure Lengend Snippet: Effects of nitroxyl hydrogen on protein expression of SERCA2a. Values are means ± SD, *P<0.01 vs. sham. ΔP<0.05 vs. HF.

Article Snippet: Angeli’s salt (Cayman), rat amino-terminal forebrain natriuretic peptide ELISA kit (Abnova), cell lysate, protease inhibitor, BCA protein concentration assay kit (all from Wuhan Boster), SERCA2a primary antibody (Abnova), goat anti-mouse secondary antibody (Abnova), β-actin primary antibody (Abnova), nitrocellulose membrane (Abnova), ATPase assay kit (Nanjing Jiancheng Institute of Bioengineering).

Techniques: Expressing