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I H Segel

Publications and source records attributed to I H Segel.

At least 73 records · Page 4Linked to original sources

Regulation of Nitrate Uptake in Penicillium chrysogenum by Ammonium Ion.

A nitrate uptake system is induced (along with nitrate reductase) when NH(4) (+)-grown Penicillium chrysogenum is incubated with inorganic nitrate in synthetic medium in the absence of NH(4) (+). Nitrate uptake and nitrate reduction are probably in steady state in fully induced mycelium, but the ratios of the two activities are not constant during the induction period. Substrate concentrations of ammonium cause a rapid decay of nitrate uptake and nitrate reductase activity. The two activities are differentially inactivated (the uptake activity being more sensitive). Glutamine and asparagine are as effective as NH(4) (+) in suppressing nitrate uptake activity. Glutamate and alanine were about half as effective as NH(4) (+). Cycloheximide interferes with the NH(4) (+)-induced decay of nitrate uptake activity. The ammonium transport system is almost maximally deinhibited (or derepressed) in nitrate-grown mycelium.

Journal Article↗

Regulation of nigeran accumulation by Aspergillus aculeatus.

Nigeran (mycodextran), an unbranched glucan consisting of alternating alpha,1-4 and alpha,1-3 linkages, is accumulated intracellularly by Aspergillus aculeatus, under conditions of nitrogen limitation and low pH (optimum pH 5.0). Both conditions are necessary. Nigeran is synthesized de novo from exogenous glucose and accounts for about 20% of the glucose transported by the mycelium. The polymer is diluted out but not degraded when the mycelium is transferred to fresh medium containing NH(4) (+). Cycloheximide prevents development of the nigeran biosynthetic system. The addition of substrate levels of NH(4) (+) to nigeran-accumulating mycelium immediately suppresses further nigeran buildup, presumably by diverting glucose to the biosynthesis of amino acids, nucleotides, etc.

Ammonium Chloride↗

Effect of weak acids on amino acid transport by Penicillium chrysogenum: evidence for a proton or charge gradient as the driving force.

A variety of weak acids at and below their pK(a) are potent inhibitors of transport in Penicillium chrysogenum. The effective compounds include sorbate, benzoate, and propionate (common antifungal agents), indoleacetate (a plant hormone), acetylsalicylate (aspirin), hexachlorophene, and a yellow pigment produced by the mycelia under nutrient-deficient conditions, as well as the classical uncouplers 2,4-dinitrophenol, p-nitrophenol, and azide. The results suggest that a proton gradient or charge gradient is involved in energizing membrane transport in P. chrysogenum. The unionized form of the weak acids could discharge the gradient by diffusing through the membrane and ionizing when they reach an interior compartment of higher pH. Experiments with 2,4-dinitrophenol and p-nitrophenol established that the ionized species are not absorbed by the mycelium to any great extent. The transport inhibitors also caused a decrease in cellular adenosine 5'-triphosphate (ATP) levels, but there was no constant correlation between inhibition of transport and suppression of cellular ATP. A decrease in aeration of the mycelial suspension had the same effect on transport and ATP levels as the addition of a weak organic acid. The effects on transport rates and ATP levels were reversible. The instantaneous inhibition of [(14)C]l-leucine transport by NH(4) (-) (and vice-versa) in nitrogen-starved mycelia at pH values of 7 or below can be explained by competition for a common energy-coupling system. The inhibition is not observed in carbon-starved mycelia in which the NH(4) (+) transport system is absent or inactive (but the general amino acid transport is fully active), or in iodoacetate-treated mycelia in which the NH(4) (+) transport system has been differentially inactivated. At pH values greater than 7.0, NH(3) and HPO(4) (2-) inhibit transport, presumably by discharging the membrane proton or charge gradient. Aniline counteracts the inhibitory effect of NH(3) and HPO(4) (2-) possibly by acting as a proton reservoir or buffer within the membrane.

Acids↗

Effect of cycloheximide on L-leucine transport by Penicillium chrysogenum: involvement of calcium.

Cycloheximide (actidione) has an immediate inhibitory effect on amino acid transport by nitrogen-starved or carbon-starved mycelium suspended in phosphate buffer. High concentrations of phosphate alone are slightly inhibitory; cycloheximide appears to potentiate the effect of phosphate. Ca(2+) reverses the inhibition of transport caused by phosphate plus cycloheximide. Ca(2+) did not relieve the inhibition of protein synthesis. Cycloheximide promotes a continual uptake of (45)Ca(2+) by the mycelium. The cumulative results suggest that (i) membrane-bound Ca(2+) is involved in amino acid transport, (ii) cycloheximide labilizes the membrane-bound Ca(2+), and (iii) phosphate forms a complex with Ca(2+) making it unavailable for its role in transport. The effect of cycloheximide described above is observed within 1 to 2 min after addition of the antibiotic. This initial inhibition occurs more rapidly with 10(-3) M cycloheximide than with 10(-5) M cycloheximide. However, after a longer preincubation time, a curious inverse relationship between cycloheximide concentration and amino acid transport is observed. The mycelium incubated with 10(-5) M cycloheximide remains strongly inhibited (unless the antibiotic is washed away). The mycelium incubated with 10(-3) M cycloheximide recovers about 40% of the transport activity lost during the rapid initial phase. We have no obvious explanation for the inverse effect.

Buffers↗

Regulation of sulfate transport in filamentous fungi.

Inorganic sulfate enters the mycelia of Aspergillus nidulans, Penicillium chrysogenum, and Penicillium notatum by a temperature-, energy-, pH-, ionic strength-, and concentration-dependent transport system ("permease"). Transport is unidirectional. In the presence of excess external sulfate, ATP sulfurylase-negative mutants will accumulate inorganic sulfate intracellularly to a level of about 0.04 m. The intracellular sulfate can be retained against a concentration gradient. Retention is not energy-dependent, nor is there any exchange between intracellular (accumulated) and extracellular sulfate. The sulfate permease is under metabolic control. Sulfur starvation of high methionine-grown mycelia results in about a 1000-fold increase in the specific sulfate transport activity at low external sulfate concentrations. l-Methionine is a metabolic repressor of the sulfate permease, while intracellular sulfate and possibly l-cysteine (or a derivative of l-cysteine) are feedback inhibitors. Sulfate transport follows hyperbolic saturation kinetics with a Michaelis constant (Km) value of 6 x 10(-5) to 10(-4)m and a V(max) (for maximally sulfurstarved mycelia) of about 5 micromoles per gram per minute. Refeeding sulfur-starved mycelia with sulfate or cysteine results in about a 10-fold decrease in the V(max) value with no marked change in the Km. Azide and dinitrophenol also reduce the V(max.).

Journal Article↗