Ca(2+ )binding to complement-type repeat domains 5 and 6 from the low-density lipoprotein receptor-related protein

BACKGROUND: The binding of ligands to clusters of complement-type repeat (CR)-domains in proteins of the low-density lipoprotein receptor (LDLR) family is dependent on Ca(2+ )ions. One reason for this cation requirement was identified from the crystal structure data for a CR-domain from the prototypic LDLR, which showed the burial of a Ca(2+ )ion as a necessity for correct folding and stabilization of this protein module. Additional Ca(2+ )binding data to other CR-domains from both LDLR and the LDLR-related protein (LRP) have suggested the presence of a conserved Ca(2+ )cage within CR-domains from this family of receptors that function in endocytosis and signalling. RESULTS: We have previously described the binding of several ligands to a fragment comprising the fifth and the sixth CR-domain (CR56) from LRP, as well as qualitatively described the binding of Ca(2+ )ions to this CR-domain pair. In the present study we have applied the rate dialysis method to measure the affinity for Ca(2+), and show that CR56 binds 2 Ca(2+ )ions with an average affinity of K(D )= 10.6 microM, and there is no indication of additional Ca(2+ )binding sites within this receptor fragment. CONCLUSIONS: Both CR-domains of CR56 bind a single Ca(2+ )ion with an affinity of 10.6 microM within the range of affinities demonstrated for several other CR-domains

Effect of implantable cardioverter-defibrillators in patients with non-ischaemic systolic heart failure and concurrent coronary atherosclerosis

AIMS: Prophylactic implantable cardioverter‐defibrillators (ICD) reduce mortality in patients with ischaemic heart failure (HF), whereas the effect of ICD in patients with non‐ischaemic HF is less clear. We aimed to investigate the association between concomitant coronary atherosclerosis and mortality in patients with non‐ischaemic HF and the effect of ICD implantation in these patients. METHODS AND RESULTS: Patients were included from DANISH (Danish Study to Assess the Efficacy of Implantable Cardioverter Defibrillators in Patients with Non‐Ischaemic Systolic Heart Failure on Mortality), randomizing patients to ICD or control. Study inclusion criteria for HF were left ventricular ejection fraction ≤ 35% and increased levels (>200 pg/mL) of N‐terminal pro‐brain natriuretic peptide. Of the 1116 patients from DANISH, 838 (75%) patients had available data from coronary angiogram and were included in this subgroup analysis. We used Cox regression to assess the relationship between coronary atherosclerosis and mortality and the effect of ICD implantation. Of the included patients, 266 (32%) had coronary atherosclerosis. Of these, 216 (81%) had atherosclerosis without significant stenoses, and 50 (19%) had significant stenosis. Patients with atherosclerosis were significantly older {67 [interquartile range (IQR) 61–73] vs. 61 [IQR 54–68] years; P < 0.0001}, and more were men (77% vs. 70%; P = 0.03). During a median follow‐up of 64.3 months (IQR 47–82), 174 (21%) of the patients died. The effect of ICD on all‐cause mortality was not modified by coronary atherosclerosis [hazard ratio (HR) 0.94; 0.58–1.52; P = 0.79 vs. HR 0.82; 0.56–1.20; P = 0.30], P for interaction = 0.67. In univariable analysis, coronary atherosclerosis was a significant predictor of all‐cause mortality [HR, 1.41; 95% confidence interval (CI), 1.04–1.91; P = 0.03]. However, this association disappeared when adjusting for cardiovascular risk factors (age, gender, diabetes, hypertension, smoking, and estimated glomerular filtration rate) (HR 1.05, 0.76–1.45, P = 0.76). CONCLUSIONS: In patients with non‐ischaemic systolic heart failure, ICD implantation did not reduce all‐cause mortality in patients either with or without concomitant coronary atherosclerosis. The concomitant presence of coronary atherosclerosis was associated with increased mortality. However, this association was explained by other risk factors