Abstract. Objectives . Patients with iron deficiency anaemia complain of decreased exercise capacity. We asked whether this is due to defective oxidative ATP synthesis in skeletal muscle as a consequence of reduced blood oxygen content and/or intrinsic mitochondrial abnormalities. Design . We used 31 P magnetic resonance spectroscopy to examine skeletal muscle bioenergetics in iron‐deficient patients and in age‐ and sex‐matched controls. Setting . The patients were recruited from the primary care population. Subjects . We studied seven symptomatic female iron‐deficient patients (aged 32–70 years) with haemoglobin (Hb) concentration, [Hb], 8.0 g dl −1 . Six had menorrhagia, the cause in the seventh patient remained undiagnosed. Results were compared with those of 8 healthy female controls (aged 25–48 years) with mean [Hb] 13.7 g dl −1 . Results . The right calf muscle was by studied 31 P magnetic resonance spectroscopy in a 1.9 T superconducting magnet. We measured the intracellular concentrations of phosphocreatine (PCr), inorganic phosphate (Pi), adenosine triphosphate (ATP) and the intracellular pH at rest, during plantar flexion exercise and during recovery from exercise. Exercise duration was reduced in the patients, yet end‐exercise PCr/(PCr + Pi) was higher and adenosine diphosphate (ADP) lower than in controls. After exercise, initial PCr recovery was slowed but this was probably because of the lower cytosolic ADP concentration. Conclusions . Mitochondrial ATP synthesis was not limited by oxygen supply or an intrinsic mitochondrial defect. Therefore, the reduced exercise capacity seen in iron deficiency could be due to central causes and not to skeletal muscle metabolic abnormalities.
Rats were fed a diet containing 1% beta-guanidino-propionic acid (GPA) for 6-12 wk to deplete their muscles of phosphocreatine (PCr). Gated 31P nuclear magnetic resonance (NMR) spectra were obtained from the gastrocnemius-plantaris muscle at various time points during either a 1- or 3-s isometric tetanic contraction using a surface coil. The energy cost of a 1-s tetanus in unfatigued control rat muscle was 48.4 mumol ATP X g dry wt-1 X s-1 and was largely supplied by PCr; anaerobic glycogenolysis was negligible. In GPA-fed rats PCr was undetectable after 400 ms. This had no effect on initial force generated per gram, which was not significantly different from controls. Developed tension in a 3-s tetanus in GPA-fed rats could be divided into a peak phase (duration 0.8-0.9 s) and a plateau phase (65% peak tension) in which PCr was undetectable and the [ATP] was less than 20% of that in control muscle. Energy from glycogenolysis was sufficient to maintain force generation at this submaximal level. Mean net glycogen utilization per 3-s tetanus was 78% greater than in control muscle. However, the observed decrease in intracellular pH was less than that expected from energy budget calculations, suggesting either increased buffering capacity or modulation of ATP hydrolysis in the muscles of GPA-fed rats. Our results demonstrate that the transport role of PCr is not essential in contracting muscle in GPA-fed rats. PCr is probably important in this regard in the larger fibers of control muscle. Although fast-twitch muscles depleted of PCr have nearly twice the glycogen reserves of control muscle, glycogenolysis is limited in its capacity to fill the role of PCr as an energy buffer under conditions of maximum ATP turnover.
1. Modification of a single amino acid residue by introduction of the nitrobenzofurazan group inactivates mitochondrial ATPase (adenosine triphosphatase) when membrane-bound in submitochondrial particles. The similarity between the reactions of both membrane-bound and isolated ATPase with 4-chloro-7-nitrobenzofurazan indicates that the single essential tryosine residue identified in the isolated enzyme [Ferguson, Loyd, Lyons & Radda (1975) Eur. J. Biochem. 54, 117-126] Is also a feature of the membrane-bound ATPase. 2. A procedure is presented for estimating the ATPase content of the inner mitochondrial membrane. It is based on the specificity of the incorporation of the nitrobenzofurazan group, and the ready removal of this group by compounds that contain a thiol group. This method indicates that 8.5% of the membrane protein is ATPase. The procedure should be applicable to the titration of the energy-transducing ATPases of bacterial plasma membranes and of the thylakoid membranes of chloroplasts. 3. Combination of the data obtained on the ATPase content of the bovine heart inner mitochondrial membrane with a titration of the cytochrome bc1 complex with antimycin indicates that these two components of the membrane are present in approximately equal amounts.
High-resolution 31P nuclear magnetic resonance spectra at 73.83 MHz are reported for rat heart in vivo. In live rats, it was possible to observe the cardiac content of ATP, phosphocreatine, and Pi. Only a small amount of whole-blood 2,3-diphosphoglycerate was observed in the spectra, precluding the possibility that blood phosphate compounds were masking the spectra of cardiac phosphate compounds. The 31P nuclear magnetic resonance spectra of in vivo and perfused rat hearts were similar and support the utilization of the perfused rat heart as a model system for studying high-energy phosphate metabolism of the heart in vivo. The dynamic flux of high-energy phosphate compounds was investigated by subjecting the rat to respiratory arrest. In this experiment, the heart followed the classic metabolic pattern known to occur during cardiac arrest; phosphocreatine and then ATP decreased in concentration while Pi increased in concentration. The 31P nuclear magnetic resonance analysis of rat heart in vivo is demonstrated to be a practical and feasible method for studying cardiac high-energy phosphate metabolism.
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