Annotating proteins from endoplasmic reticulum and Golgi apparatus in eukaryotic proteomes

The sub-cellular localization of a native protein constitutes one coarse-grained aspect of its function. Transport between compartments is often regulated through short sequence motifs. Here, we analyzed experimentally characterized endoplasmic reticulum (ER)/ Golgi retrieval motifs and investigated the accuracy of homology-transfer. Only the C-terminal ER retrieval motifs KDEL, HDEL and AIAKE were sufficiently specific. However, even unspecific motifs may help, provided we know the probability for localization given the motif. We provided such estimates. We also rigorously estimated the accuracy and coverage for inferring ER and Golgi localization through homology-transfer by sequence similarity. In entire proteomes, we could thereby annotate 3304 ER (3182 membrane) and 1853 Golgi (759 membrane) proteins. We identified another putative 5157 globular and 3941 membrane ER or Golgi proteins. Each experimental annotation yielded, on average, one to three high-accuracy and five to six low-accuracy homology-transfers in the six proteomes. These numbers will increase with each new experimental annotation.Received 6 January 2004; received after revision 10 March 2004; accepted 29 March 2004     

The effect of adrenaline on the electrogenic Na+ pump in cardiac muscle cells

Electrogenic Na+ pump currents during K+-activated hyperpolarizations of bullfrog atrium muscle fibres are increased by adrenaline. The log dose-response relation between these currents and activating K+ concentrations is expressed by a sigmoidal curve, which is shifted in parallel to the left by adrenaline. It is suggested that adrenaline increases the rate of Na+ extrusion without increasing the Na/K coupling ratio and total number of pumping sites.     

Direct toxic effect of isoproterenol on cultured cardiac muscle cells

Summary Isoproterenol at relatively high doses (2.5 mg/ml) has a marked toxic effect on rat heart muscle cells cultivated in vitro. This effect is not prevented by propranolol and therefore is not mediated by beta adrenergic receptors.This work has been supported in part by a grant from the Muscular Dystrophy Association, Inc. to Prof. M. Aloisi.     

Localization of pyruvate carboxylase in the cells ofNeurospora crassa

Summary The cell wall ofNeurospora crassa was digested enzymatically and the cytosolic and the mitochondrial fractions were separated. The activity of pyruvate carboxylase (EC 6.4.1.1) was detected entirely in the cytosolic fraction. This indicates that the location of pyruvate carboxylase ofN. crassa is in the cytosol, but is not in the mitochondria; this is different from the situation in animal tissues.     

Localization of pyruvate carboxylase in the cells of Neurospora crassa

The cell wall of Neurospora crassa was digested enzymatically and the cytosolic and the mitochondrial fractions were separated. The activity of pyruvate carboxylase (EC 6.4.1.1) was detected entirely in the cytosolic fraction. This indicates that the location of pyruvate carboxylase of N. crassa is in the cytosol, but is not in the mitochondria; this is different from the situation in animal tissues.