During inhibition of cell growth by weak acids, there is substantial accumulation of the weak acid anions in the cytoplasm. This study was undertaken to determine the impact of anion accumulation on cellular pools. At pH 6, growth in the presence of 8 mM acetate led to an internal pool of greater than 240 mM acetate anion and resulted in reduced levels of glutamate in the cell, but there were no significant changes in K+ and Na+ levels. At low osmolarity, the change in the glutamate pool compensated for only a small fraction of the accumulated acetate anion. However, at high osmolarity, glutamate compensated for over half of the accumulated acetate. Recovery of the normal cytoplasmic pH after the removal of acetate was dependent on the synthesis of glutamate.

Cadmium-induced metallothioneins from the common sea mussel, Mytilus edulis, were shown to comprise of two groups of isoforms having apparent molecular masses of 10kDa and 20kDa. The 1 0-kDa group was resolved by anion-exchange chromatography into four fractions while the 20-kDa group was resolved into three fractions using this method. After metal removal and Smethylation of the cysteine residues using methyl-p-nitrobenzenesulphonate the complete amino acid sequences were determined. Five isoforms of the 20-kDa group were shown to possess monomeric units consisting of 71 amino acids. These proteins were distinct from the four 72-amino-acid proteins of the 10-kDa group. The FASTA algorithm has been used to compare the degree of similarity between the mussel metallothionein MT-10-IV isoform and other metallothioneins. The mussel MT-10-IV isoform exhibited substantial similarity to other molluscan metallothioneins. Moreover, the mussel metallothionein exhibited more similarity to vertebrate metallothioneins than to those of non-molluscan invertebrates, thus suggesting that the mussel metallothioneins are class I metallothioneins.Despite the discovery of the small cysteine-rich metalbinding-protein metallothionein (MT) more than 30 years ago in equine kidney cortex, the function of this protein is still not fully understood. MT is known to bind metals such as Zn and Cu (which are essential for cell growth and development) and Cd and Hg (which are toxic), which suggests that MT might have more than one function.The search for putative functions for these proteins is exacerbated by the existence of numerous inducers and isoforms. The range of factors which regulate MT synthesis and degradation together with the ubiquity and sequence conservation of MT perhaps suggests that these proteins have a central role in cell metabolism. The proposed functions for MT include the detoxification of heavy metals and the metabolism of essential metals. Zeng et al. (1991) have suggested that thionein may be the active form of the protein and it may be involved in the control of gene expression by virtue of its ability to remove Zn from Zn-finger proteins. Moreover, a tissue-specific isoform of MT in astrocytes (also called growth-inhibitory factor) which inhibits the survival Correspondence to

No abstract

Atlantic cod, Gadus morhua, were maintained on a diet of sandeel and after a 6-day fast were refed a single meal. Concentrations of free amino acids (AAs) were measured in hepatic portal and cardiac blood as well as in the stomach and white muscle at intervals of 6h up to 24h post-feeding. The appearance of both essential and non-essential AAs in the hepatic portal blood was significantly correlated, up to 12h after feeding, to their abundance in the diet. There was a significant decline in total AA concentration in cardiac blood after 6h, followed by a significant increase at 12h. No significant changes in total AA concentration were observed in the other tissues, although mean concentration increased at 12 or 18h. At a more detailed level, the post-prandial changes in concentration of some essential AAs were consistent with their having a role in the stimulation of protein synthesis after feeding.

The induced uptakes of L-[3H]phenylalanine and L-[3H]arginine in oocytes injected with clonal NBAT (neutral and basic amino acid transporter) cRNA show differential inactivation by pretreatment with N-ethylmaleimide (NEM), revealing at least two distinct transport processes. NEM-resistant arginine transport is inhibited by leucine and phenylalanine but not by alanine or valine; mutual competitive inhibition of NEM-resistant uptake of arginine and phenylalanine indicates that the two amino acids share a single transporter. NEM-sensitive arginine transport is inhibited by leucine, phenylalanine, alanine and valine. At least two NEM-sensitive transporters may be expressed because we have been unable to confirm mutual competitive inhibition between arginine and phenylalanine transport. The NEM-resistant transport mechanism appears to involve distinct but overlapping binding sites for cationic and zwitterionic substrates. NBAT is known to form oligomeric protein complexes in cell membranes, and its functional roles when expressed in Xenopus oocytes may include interaction with oocyte proteins, leading to increased native amino acid transport activities; these resemble NBAT-expressed activities in terms of NEM-sensitivity and apparent substrate range (including an unusual inhibition by beta-phenylalanine.