Hsp40 chaperones promote degradation of the HERG potassium channel.

Loss of function mutations in the hERG (human ether-a-go-go related gene or KCNH2) potassium channel underlie the proarrhythmic cardiac long QT syndrome type 2. Most often this is a consequence of defective trafficking of hERG mutants to the cell surface, with channel retention and degradation at the endoplasmic reticulum. Here, we identify the Hsp40 type 1 chaperones DJA1 (DNAJA1/Hdj2) and DJA2 (DNAJA2) as key modulators of hERG degradation. Overexpression of the DJAs reduces hERG trafficking efficiency, an effect eliminated by the proteasomal inhibitor lactacystin or with DJA mutants lacking their J domains essential for Hsc70/Hsp70 activation. Both DJA1 and DJA2 cause a decrease in the amount of hERG complexed with Hsc70, indicating a preferential degradation of the complex. Similar effects were observed with the E3 ubiquitin ligase CHIP. Both the DJAs and CHIP reduce hERG stability and act differentially on folding intermediates of hERG and the disease-related trafficking mutant G601S. We propose a novel role for the DJA proteins in regulating degradation and suggest that they act at a critical point in secretory pathway quality control.

Co-chaperone FKBP38 promotes HERG trafficking.

The Long QT Syndrome is a cardiac disorder associated with ventricular arrhythmias that can lead to syncope and sudden death. One prominent form of the Long QT syndrome has been linked to mutations in the HERG gene (KCNH2) that encodes the voltage-dependent delayed rectifier potassium channel (I(Kr)). In order to search for HERG-interacting proteins important for HERG maturation and trafficking, we conducted a proteomics screen using myc-tagged HERG transfected into cardiac (HL-1) and non-cardiac (human embryonic kidney 293) cell lines. A partial list of putative HERG-interacting proteins includes several known components of the cytosolic chaperone system, including Hsc70 (70-kDa heat shock cognate protein), Hsp90 (90-kDa heat shock protein), Hdj-2, Hop (Hsp-organizing protein), and Bag-2 (BCL-associated athanogene 2). In addition, two membrane-integrated proteins were identified, calnexin and FKBP38 (38-kDa FK506-binding protein, FKBP8). We show that FKBP38 immunoprecipitates and co-localizes with HERG in our cellular system. Importantly, small interfering RNA knock down of FKBP38 causes a reduction of HERG trafficking, and overexpression of FKBP38 is able to partially rescue the LQT2 trafficking mutant F805C. We propose that FKBP38 is a co-chaperone of HERG and contributes via the Hsc70/Hsp90 chaperone system to the trafficking of wild type and mutant HERG potassium channels.

Cardiopulmonary physiology and responses of ultramarathon athletes to prolonged exercise.

The purpose of this study was to determine the changes of pulmonary function and autonomic cardiovascular control after an ultramarathon and their relation to performance. Eight entrants to the Canadian National Championship 100-km running race participated in the study. Pulmonary function and 30-s maximum voluntary ventilation (MVV30s) tests were conducted one day before the race and within 5 minutes of race completion. Heart rate and blood pressure data were collected 30 min before and 5 min after the race as well as during a 10-min stand test one day prior to the race. During the race, beat-by-beat R-R interval data were collected over the first and last 20 km. The results showed that MVV30s and MVV30s tidal volumes were reduced postrace (p < 0.001). Prerace supine total harmonic variation (p < 0.01) and prerace MVV values (10 s to 30 s) (p < 0.05) were correlated with race finish time. The changes in pulmonary function and MVV30s values from pre- and postrace were not significantly correlated to race performance. We conclude that maximal sustainable ventilatory power and dynamic autonomic cardiovascular control are important factors in determining overall performance in an ultramarathon.