Structural and functional implications of the phospholamban hinge domain: impaired SR Ca2+ uptake as a primary cause of heart failure.
OBJECTIVE
The role of sarcoplasmic reticulum (SR) in the onset and progression of heart failure is controversial. We tested the hypothesis that impairment of SR Ca2+ sequestration may be a primary cause for progressive left ventricular (LV) dysfunction and the phospholamban hinge domain may be critical in this process.


METHODS
A phospholamban hinge domain mutant (PLB/N27A) was introduced in the cardiac compartment of the phospholamban null mouse. An integrative approach was used to characterize the resulting cardiac phenotype at a structural, cellular, whole organ and intact animal level.


RESULTS
NMR analysis revealed a defined alteration in the alpha-helical configuration between residues Q22 to F35 in mutant phospholamban. Transgenic lines expressing similar levels of mutant compared to wild-type phospholamban exhibited super-inhibition of the SR Ca2+ ATPase affinity for Ca2+ (EC50 0.52 microM) in oxalate-supported Ca2+ uptake measurements, which translated into impaired relaxation and attenuated responses to beta-adrenergic stimulation. Importantly, a blunted force-frequency relation was observed in mutant hearts preceding left ventricular dilation. Upon aging to 10 months, the predominantly diastolic dysfunction progressed to congestive heart failure, characterized by induction of a fetal gene program, cardiac remodeling, lung congestion, depressed systolic function and early mortality.


CONCLUSIONS
Increased inhibition of Ca2+ sequestration may be a causative factor in the development of left ventricular dysfunction and myocyte remodeling leading to heart failure. Furthermore, the hinge domain may play an important role in transmitting PLB's regulatory effects on SERCA.
Schmidt
Albrecht G
AG
Department of Pharmacology and Cell Biophysics, University of Cincinnati College of Medicine, 231 Albert Sabin Way, Cincinnati, OH 45267-0575, USA.
Zhai
Jing
J
Carr
Andrew N
AN
Gerst
Mike J
MJ
Lorenz
John N
JN
Pollesello
Piero
P
Annila
Arto
A
Hoit
Brian D
BD
Kranias
Evangelia G
EG
D000375:Aging / Q000502:physiology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D018450:Disease Progression; D006333:Heart Failure / Q000378:metabolism* / Q000473:pathology; D006439:Hemodynamics; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009154:Mutation; D009200:Myocardial Contraction; D017433:Protein Structure, Secondary; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*; D013329:Structure-Activity Relationship; D015996:Survival Rate; D018487:Ventricular Dysfunction, Left / Q000378:metabolism; D000099211:Phospholamban
D002118:Calcium; D002135:Calcium-Binding Proteins; D000099211:Phospholamban