TY  - JOUR
U1  - Zeitschriftenartikel, wissenschaftlich - begutachtet (reviewed)
A1  - Herwig, Melissa
A1  - Kolijn, Detmar
A1  - Lódi, Mária
A1  - Hölper, Soraya
A1  - Kovács, Árpád
A1  - Papp, Zoltán
A1  - Jaquet, Kornelia
A1  - Haldenwang, Peter
A1  - Dos Remedios, Cris
A1  - Reusch, Peter H.
A1  - Mügge, Andreas
A1  - Krüger, Marcus
A1  - Fielitz, Jens
A1  - Linke, Wolfgang A.
A1  - Hamdani, Nazha
T1  - Modulation of Titin-Based Stiffness in Hypertrophic Cardiomyopathy via Protein Kinase D
JF  - Frontiers in Physiology
N2  - The giant protein titin performs structure-preserving functions in the sarcomere and is important for the passive stiffness (Fpassive) of cardiomyocytes. Protein kinase D (PKD) enzymes play crucial roles in regulating myocardial contraction, hypertrophy, and remodeling. PKD phosphorylates myofilament proteins, but it is not known whether the giant protein titin is also a PKD substrate. Here, we aimed to determine whether PKD phosphorylates titin and thereby modulates cardiomyocyte Fpassive in normal and failing myocardium. The phosphorylation of titin was assessed in cardiomyocyte-specific PKD knock-out mice (cKO) and human hearts using immunoblotting with a phosphoserine/threonine and a phosphosite-specific titin antibody. PKD-dependent site-specific titin phosphorylation in vivo was quantified by mass spectrometry using stable isotope labeling by amino acids in cell culture (SILAC) of SILAC-labeled mouse heart protein lysates that were mixed with lysates isolated from hearts of either wild-type control (WT) or cKO mice. Fpassive of single permeabilized cardiomyocytes was recorded before and after PKD and HSP27 administration. All-titin phosphorylation was reduced in cKO compared to WT hearts. Multiple conserved PKD-dependent phosphosites were identified within the Z-disk, A-band and M-band regions of titin by quantitative mass spectrometry, and many PKD-dependent phosphosites detected in the elastic titin I-band region were significantly decreased in cKO. Analysis of titin site-specific phosphorylation showed unaltered or upregulated phosphorylation in cKO compared to matched WT hearts. Fpassive was elevated in cKO compared to WT cardiomyocytes and PKD administration lowered Fpassive of WT and cKO cardiomyocytes. Cardiomyocytes from hypertrophic cardiomyopathy (HCM) patients showed higher Fpassive compared to control hearts and significantly lower Fpassive after PKD treatment. In addition, we found higher phosphorylation at CaMKII-dependent titin sites in HCM compared to control hearts. Expression and phosphorylation of HSP27, a substrate of PKD, were elevated in HCM hearts, which was associated with increased PKD expression and phosphorylation. The relocalization of HSP27 in HCM away from the sarcomeric Z-disk and I-band suggested that HSP27 failed to exert its protective action on titin extensibility. This protection could, however, be restored by administration of HSP27, which significantly reduced Fpassive in HCM cardiomyocytes. These findings establish a previously unknown role for PKDin regulating diastolic passive properties of healthy and diseased hearts.
KW  - -
KW  - titin
KW  - HCM
KW  - PKD
KW  - Hsp27
KW  - stiffness
Y1  - 2020
UN  - https://nbn-resolving.org/urn:nbn:de:gbv:9-opus-36353
SN  - 1664-042X
SS  - 1664-042X
U6  - https://doi.org/10.3389/fphys.2020.00240
DO  - https://doi.org/10.3389/fphys.2020.00240
VL  - 11
PB  - Frontiers Media S.A.
ER  -