Biomedical subjects

A Kotyk

Publications and source records attributed to A Kotyk.

At least 109 records · Page 6Linked to original sources

PubMed1976

Monosaccharide transport systems in the yeast Rhodotorula glutinis.

By using D-glucose, D-xylose, D-galactose and D-fructose in the strictly aerobic yeast Rhodotorula glutinis and by comparing the half-saturation constants with inhibition constants the yeast was shown to possess a single common system for D-xylose and D-galactose (Km's and Ki's all between 0.5 and 1.1 mM) but another distinct transport system for D-fructose. The transport of D-glucose has a special position in that glucose blocks apparently allotopically all the other systems observed although it uses at least one of them for its own transport. The different character of D-glucose uptake is underlined by its relative independence of pH (its "Km" is completely pH-insensitive) in contrast with all other sugars. At low concentrations, all sugars show mutual positive cooperativity in uptake, suggesting at least two transport sites plus possibly a modifier site on the carrier.

Binding Sites↗

PubMed1976

Some properties of the adenosine triphosphatase systems of two yeast species, Saccharomyces cerevisiae and Rhodotorula glutinis.

1. Total ATPase levels were determined in homogenate fractions of baker's yeast, Saccharomyces cerevisiae K and Rhodotorula glutinis. The maximum ATPase activities in 8000 X g supernatant of the three yeast strains were 6.0, 1.9, and 2.2 mmol Pih-1 (gDS)-1, respectively; the activities in the sediment were somewhat higher. Exponential cells of S. cerevisiae K and R. glutinis exhibited higher ATPase levels than did the stationary cells. 2. The total ATPase activity in both yeast species showed a maximum at ph 6.8 a minimum at pH 7.2, and another broader masimum around pH 8.0. 3. No significant NaK-ATPase activity was detected in baker's yeast, in either the exponential or the stationary cells of R. glutinis, and in exponential S. cerevisiae K cells in the pH range of 6.0-9.3. 4. Stationary cells of S. cerevisiae K exhibited, at pH 7.0-8.5, A Na,K-ATPase activity attaining 9% of total ATPase level. 5.3 X 10(-3) M phenylmethyl sulphonyl fluoride had no effect on the total ATPase level in S. cerevisiae and inhibited the activity in R. glutinis by 25%; it did not bring forth any Na,K-ATPase activity apart from that found in its absence. 6. 1.5 M urea lowered the ATPase activity in R. glutinis by 68% but had no effect on S. cerevisiae cells. 10(-5) M dicyclohexylcarbodiimide suppressed the ATPase activity in S. cerevisiae and R. glutinis by 74 and 79%, respectively. Neither agent revealed and additional Na,K-ATPase activity. 7. The comparison of Na,K-ATPase activities with data on K+ fluxes across the yeast plasma membrane suggested that even with the lower flux values the Na,K-ATPase, even if present, would account for a mere 40% of transported ions. The results imply that the active ion transport in yeasts is energized by mechanisms other than the Na,K-ATPase.

Adenosine Triphosphatases↗

PubMed1976

Loss of inducible D-galactose transport by baker's yeast after osmotic treatment.

The ability of Saccharomyces cerevisiae to transport D-galactose and related sugars with an axial hydroxyl group at C-4, acquired by induction with D-galactose, was lost either by exposing early exponential-phase cells to an osmotic shock involving incubation in 0.6M NaC1O4, 0.66M sucrose and 1mM histidine and transfer to 5mM Tris-HC1 with 2mM dithiothreitol, or simply by transferring them to distilled water. The total amount of protein thus released was 0.1--0.35 and 0.1 mg per mg dry wt., respectively. The shock fluid contained at least six proteins, among them a galactose-binding component. L-Arabinose transport could not be restored by adding the concentrated shock fluid to depleted cells but cells remained viable after the shock and resynthesized the transport system if incubated in a galactose-containing growth medium.

Arabinose↗

PubMed1976

Uptake and binding of dissaccharides in human erythrocytes.

Disaccharides (sucrose, lactose, melibiose, cellobiose, trehalose, maltose, and isomaltose) are not transported across the human erythrocyte membrane. Maltose alone is bound in appreciable amounts to the intact cell as well as ghost membranes and competes mutually for uptake with D-glucose. In (NH4)2-SO4-precipitated membrane preparations, maltose binds more strongly than other disaccharides (KD = 1.3 X 10(-5) M; maximum binding capacity, 71 pmol/mg protein) and again competes mutually with D-glucose. Phloretin inhibits the binding of glucose much more than that of maltose.

Binding Sites↗

PubMed1975

Isolation and properties of an arginine-binding protein from Saccharomyces cerevisiae.

Transfer of exponentially growing cells of Saccharomyces cerevisiae epsilon 1278 b to a fresh medium (or simply to distilled water) resulted in the loss of ability to transport arginine (and lysine), accompanied by the release of several proteins from the membrane surface or periplasmic space. Fractionation by ultrafiltration, Sephadex G-50 chromatography and freeze-drying yielded a homogeneous protein (55 mg per 100 g dry weight of cells) with specific binding ability for L-arginine (Kd = 3.8 X 10(-1) M) and L-lysine (Ki = 4.2 X 10(-4) M). The protein contains over 40 amino acid residues and has a molecular weight of about 5,000. In solution, it appears to aggregate as its concentration is raised, thereby decreasing the overall binding capacity for arginine. Addition of the protein to a depleted culture does not restore the transport of arginine. It is apparently the recognition protein for the specific arginine-transporting system of Saccharomyces cerevisiae but it occurs in almost identical amounts in the MG 168 mutant with impaired arginine transport.

Amino Acids↗

PubMed1975

Transport of acyclic polyols in Saccharomyces cerevisiae.

Acyclic polyols (erythritol, xylitol, ribitol, D-arabinitol, mannitol, sorbitol and galactitol) are not metabolized by Saccharomyces cerevisiae. They are taken up by a fast non-active process, reaching 40-70% distribution referred to total cell water. The uptake is insensitive to temperature, pH (between 4 and 8), 2,4-dinitrophenol and uranyl ions. Its initial rate rises linearly with concentration from 10(-5)M to 1M. The process resembles simple diffusion through large pores or the trapping of the whole solution on the surface. Protoplasts behave like whole cells in this respect. Only erythritol shows a second type of uptake which is inhibitor-insensitive but temperature-dependent.

Biological Transport, Active↗

PubMed1975

Transport of 4-deoxy- and 6-deoxy-D-glucose in baker's yeast.

Tritium-labelled 4-deoxy-D-glucose (4-dglc) and 6-deoxy-D-glucose (6-dgcl) were prepared by catalytic hydrogenolysis of the corresponding deoxyiodo derivatives with gaseous tritium. The two sugars are transported into Saccharomyces cerevisiae by both the constitutive glucose and the inducible galactose carrier. Uranyl ions are powerful inhibitors. The pH optimum in uninduced cells lies at 5.5 for both sugars, the apparent activation energies (between 15 and 35 degrees C) are 25.1 kJ/mol and 16.5 kJ/mol, respectively. The steady-state intracellular concentration of both sugars is less than the extracellular one (no uphill transport). Neither of them is a substrate of yeast hexokinase. 4-Deoxy-D-glucose undergoes a dinitrophenol-sensitive conversion to an unknown metabolite which is not phosphorylated and may represent one of its oxidation products.

Deoxy Sugars↗

PubMed1974