Phosphorylating Titin’s Cardiac N2B Element by ERK2 or CaMKIIδ Lowers the Single Molecule and Cardiac Muscle Force
| Autor: | Siegfried Labeit, John E. Smith, Thomas Lanzicher, Rebecca E. Slater, Henk Granzier, Brian P. Anderson, Orfeo Sbaizero, Giorgia Del Favero, John Perkin, Carlos Hidalgo |
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| Přispěvatelé: | Perkin, John, Slater, Rebecca, DEL FAVERO, Giorgia, Lanzicher, Thoma, Hidalgo, Carlo, Anderson, Brian, Smith, John E., Sbaizero, Orfeo, Labeit, Siegfried, Granzier, Henk |
| Jazyk: | angličtina |
| Předmět: |
Molecular Sequence Data
Biophysics Immunoglobulin domain Stress Sarcomere Mice medicine Myocyte Animals Humans Connectin Amino Acid Sequence Phosphorylation Mechanical Phenomena Mitogen-Activated Protein Kinase 1 biology New and Notable Kinase Animal Myocardium Cardiac muscle Force spectroscopy Mechanical Biomechanical Phenomena medicine.anatomical_structure Biochemistry biology.protein Titin Female Calcium-Calmodulin-Dependent Protein Kinase Type 2 Stress Mechanical Human |
| Zdroj: | Biophysical Journal. (12):2592-2601 |
| ISSN: | 0006-3495 |
| DOI: | 10.1016/j.bpj.2015.11.002 |
| Popis: | Titin is a large filamentous protein that is responsible for the passive force of the cardiac sarcomere. Titin's force is generated by its I-band region, which includes the cardiac-specific N2B element. The N2B element consists of three immunoglobulin domains, two small unique sequence insertions, and a large 575-residue unique sequence, the N2B-Us. Posttranslational modifications of the N2B element are thought to regulate passive force, but the underlying mechanisms are unknown. Increased passive-force levels characterize diastolic stiffening in heart-failure patients, and it is critical to understand the underlying molecular mechanisms and identify therapeutic targets. Here, we used single-molecule force spectroscopy to study the mechanical effects of the kinases calcium/calmodulin-dependent protein kinase II delta (CaMKII delta) and extracellular signal-regulated kinase 2 (ERK2) on the single-molecule mechanics of the N2B element. Both CaMKII delta and ERK2 were found to phosphorylate the N2B element, and single-molecule force spectroscopy revealed an increase in the persistence length (Lp) of the molecule, indicating that the bending rigidity of the molecule was increased. Experiments performed under oxidizing conditions and with a recombinant N2B element that had a simplified domain composition provided evidence that the Lp increase requires the N2B-Us of the N2B element. Mechanical experiments were also performed on skinned myocardium before and after phosphorylation. The results revealed a large (similar to 30%) passive force reduction caused by CaMKII delta and a much smaller (similar to 6%) reduction caused by ERK2. These findings support the notion that the important kinases ERK2 and CaMKII delta can alter the passive force of myocytes in the heart (although CaMKII delta appears to be more potent) during physiological and pathophysiological states |
| Databáze: | OpenAIRE |
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