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PubMed1985

Direct determination of leucine metabolism and protein breakdown in humans using L-[1-13C, 15N]-leucine and the forearm model.

We have used the forearm model to study protein metabolism in six normal healthy subjects in the fed state using L-[1-13C, 15N]-leucine as the substrate tracer. Deep venous and arterialized venous blood samples from the forearm were collected at 10-min intervals 2.5 h into a primed-continuous infusion of the dilabelled tracer. Arterialized venous blood was obtained using a 'hot-box' technique and forearm blood flow was measured by mercury strain-gauge plethysmography. The concentration and isotope enrichment of leucine and its metabolites, alpha-ketoisocaproic acid and CO2, in deep venous and arterialized venous blood were measured by gas chromatography-mass spectrometry and isotope ratio-mass spectrometry. The rates of leucine deamination and reamination were 388 +/- 24 (mean +/- SEM) and 330 +/- 23 nmol (100 ml)-1 min-1 respectively, whilst protein synthesis and breakdown rates were 127 +/- 11 and 87 +/- 10 nmol (100 ml)-1 min-1 respectively across the forearm in the fed state. We have demonstrated that the use of doubly labelled leucine as tracer and application of the mathematical model developed in this study, permits the comprehensive quantification of leucine kinetics including protein breakdown.

Adult↗
PubMed1991

Leucine kinetics from [2H3]- and [13C]leucine infused simultaneously by gut and vein.

In amino acid tracer kinetic studies of the fed state, ingested amino acid may be taken up during its initial transit through splanchnic tissues and thus not enter the plasma compartment where tracer is infused. To investigate this possibility, adult human subjects received simultaneous intravenous (iv) and intragastric (ig) leucine tracer infusions, first during a postabsorptive (PA) 4-h primed continuous ig infusion of L-[1-13C]-leucine and L-[5,5,5-2H3]leucine iv, followed on a separate day by a fed infusion, in which an ig infusion of a liquid formula was started 2 h before the tracer infusion and continued throughout the tracer study. Subjects were accustomed to a constant experimental diet supplying 1.5 g protein.kg-1.day-1 and 41-45 kcal.kg-1.day-1 for 7 and 12 days before the PA and fed studies, respectively. For the PA study, plasma enrichment for the ig tracer was 3.34 +/- 0.27 (SE) mol + excess and for the iv tracer it was 4.18 +/- 0.10 (P less than 0.02). Enrichments of alpha-keto-isocaproic acid (KIC) were 3.24 +/- 0.16 (ig) and 3.02 +/- 0.14 (iv), respectively [not significant (NS)]. For the fed study, plasma leucine enrichment for the ig tracer was 2.15 +/- 0.14 and for the iv tracer was 2.84 +/- 0.09 (P less than 0.02). KIC enrichments were 2.02 +/- 0.08 (ig) and 2.24 +/- 0.08 (iv), respectively (NS). In the PA study, the ratio of the plasma leucine enrichments for the ig and iv tracers was 0.80 +/- 0.06 and in the fed experiment, 0.76 +/- 0.05, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

Increased leucine flux and leucine oxidation during the luteal phase of the menstrual cycle in women.

Possible changes in protein metabolism during the menstrual cycle were examined in eight healthy women who received an intravenous infusion of L-[1-13C]leucine on time during the follicular phase of the menstrual cycle and one time during the luteal phase. Enrichment of plasma [13C]ketoisocaproate and expired 13CO2 were measured to determine leucine flux and oxidation. Continuous respiratory gas exchange measurements were made for the determination of CO2 production, O2 uptake, and energy expenditure. The day of the tests, plasma thyroid hormone concentrations were measured as well as plasma and urinary cortisol. Leucine flux was higher during the luteal than during the follicular phase (2.25 +/- 0.39 vs 2.01 +/- 0.42 mumol.kg-1.min-1; P < 0.01), and leucine oxidation was also increased during the luteal phase [0.52 +/- 0.14 vs. 0.44 +/- 0.05 mumol.kg-1.min-1 (P < 0.05) for luteal and follicular phases, respectively]. Resting energy expenditure was increased during the luteal phase compared with the follicular phase (218 +/- 22 and 199 +/- 12 kJ/h, respectively). Plasma free triiodothyronine (T3) and the ratio triiodothyronine/reverse triiodothyronine (T3/rT3) were both significantly higher during the luteal phase [7.7 +/- 0.6 vs. 7.1 +/- 0.8 and 4.65 +/- 0.80 vs. 3.93 +/- 0.70 for T3 and T3/rT3, respectively (P < 0.05 for both comparisons)]. This study shows small changes in protein metabolism during the menstrual cycle in women, with an increase in oxidative leucine metabolism during the luteal phase. The concomitant increase observed in circulating free T3 raises the possibility that fluctuations in protein metabolism and thyroid hormones throughout the menstrual cycle are causally related.

Adult↗
PubMed1992

Mutation of leucine-57 to phenylalanine in a platelet glycoprotein Ib alpha leucine tandem repeat occurring in patients with an autosomal dominant variant of Bernard-Soulier disease.

The primary sequences of the three individual glycoprotein (GP) chains, GPIb alpha, GPIb beta, and GPIX, comprising the normal platelet GPIb/IX receptor for von Willebrand factor (vWF) have recently been determined, opening the possibility for characterization of disorders of this receptor at the molecular level. The presence of a leucine tandem repeat in each of these chains is of particular interest, because such repeats may be involved in associations between polypeptide segments. We now report an autosomal dominant variant of Bernard-Soulier disease associated with the heterozygous substitution of phenylalanine for a highly conserved leucine residue within the GPIb alpha leucine tandem repeat. Affected individuals experienced a moderate bleeding tendency, thrombocytopenia, and an increased mean platelet volume. Platelet aggregation was decreased only in response to ristocetin or to asialo-vWF. The kd for 125I-vWF binding to patients platelets was significantly increased over control values at 0.5 mg/mL ristocetin, but was normal at 1.0 or 1.5 mg/mL ristocetin. While sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed an essentially normal complement of all components of the GPIb/IX complex, a minor amount of a putative proteolytic fragment was identified that migrated faster than GPIb and was immunoreactive with polyclonal anti-GPIb alpha antibody, but not with a monoclonal antibody directed against the 45-Kd amino-terminal region of GPIb alpha. However, because the great majority of patient GPIb alpha comigrates with normal GPIb alpha, the major functional abnormalities of the patient platelets are most likely a consequence of the altered structure of the nonproteolyzed protein. Full concordance within the studied family between phenotypic expression and a heterozygous single nucleotide substitution in genomic DNA coding for a phenylalanine in place of the wild-type leucine at residue 57 of the mature GPIb alpha, absence of this substitution in 266 alleles from the normal population, and the lack of any other abnormality of patient DNA throughout the entire coding sequence for GPIb alpha provide strong support that this substitution may constitute a pathologic point mutation responsible for the observed phenotypic abnormalities. While the roles that leucine tandem repeats may normally play within the GPIb/IX complex are not yet known, the perturbation of such a repeat in GPIb alpha may impair interaction with other components of the complex and/or with the binding of vWF.

Adolescent↗
PubMed1991

The BZLF1 protein of EBV has a coiled coil dimerisation domain without a heptad leucine repeat but with homology to the C/EBP leucine zipper.

The EBV transactivator protein BZLF1 can bind many sites in the EBV genome, most of which have homology to a consensus AP-1 site, the binding site for the fos/jun family of transcription factors. Here we present evidence that BZLF1 can also recognise the binding site for the CCAAT/enhancer binding protein C/EBP and that a BZLF1 binding site within the BZLF1 promoter is recognised by the C/EBP protein. Analysis of the BZLF1 DNA binding domain suggests that the BZLF1 protein binds to DNA as a dimer using sequences adjacent to a basic DNA binding motif. The BZLF1 dimerisation domain does not have a heptad repeat of leucine residues common to leucine zipper proteins but does have characteristics of a coiled coil structure, as judged by site directed mutagenesis. We therefore propose that the dimerisation domain of BZLF1 is structurally related to the coiled coil structure of leucine zippers but lacks the highly conserved leucine repeat. We show that the PZLF1 dimerisation domain has residues in common with the C/EBP leucine zipper and discuss the possible implications of this relationship.

Amino Acid Sequence↗
PubMed1996

Differential inhibition of calpain and proteasome activities by peptidyl aldehydes of di-leucine and tri-leucine.

To explore membrane-permeable synthetic inhibitors that discriminate between endogenous calpain and proteasome in cells, we examined the inhibition of profiles against calpain and proteasome in vitro and in vivo of peptidyl aldehydes possessing di-leucine and tri-leucine. The tripeptide aldehyde benzyloxycarbonyl-leucyl-leucinal (ZLLLal) strongly inhibited calpain and proteasome activities in vitro. The concentration required for 50% inhibition (IC50) of the casein-degrading activity of calpain was 1.25 microM, and the IC50s for the succinyl-leucyl-leucyl-valyl-tyrosine-4-methylcoumaryl-7-amide (Suc-LLVY-MCA)- and benzyloxycarbonyl-leucyl-leucyl-leucine-4-methylcoumaryl -7-amide (ZLLL-MCA)-degrading activities of proteasome were 850 and 100 nM, respectively. On the other hand, the synthetic dipeptide aldehyde benzyloxycarbonyl-leucyl-leucinal (ZLLal) strongly inhibited the casein degrading activity of calpain (IC50 1.20 microM), but the inhibition of proteasome was weak (IC50S for SucLLVY-MCA- and ZLLL-MCA-degrading activities were 120 and 110 microM, respectively). Thus, while calpain was inhibited by similar concentrations of ZLLal and ZLLLal, the inhibitory potencies of ZLLLal against the ZLLL-MCA- and Suc-LLVY-MCA-degrading activities in proteasome were 1,100 and 140 times stronger than those of ZLLal, respectively. To evaluate the effectiveness of these inhibitors on intracellular proteasome, the induction of neurite outgrowth in PC12 cells caused by proteasome inhibition was examined. ZLLLal and ZLLal initiated neurite outgrowth with optimal concentrations of 20 nM and 10 microM, respectively, again showing a big difference in the effective concentrations for the proteasome inhibition as in vitro. As for the effect on intracellular calpain, the concentration of ZLLLal and ZLLal required for the inhibition of the autolytic activation of calpain in rabbit erythrocytes were 100 and 100 microM or more, respectively. The almost equal inhibitory potencies of ZLLLal and ZLLal were in agreement with the inhibition of calpain in vitro. These differential effects of inhibitors against calpain and proteasome are potentially useful for identifying the functions of calpain and proteasome in cell physiology and pathology.

Animals↗
PubMedDate not supplied

Comparison of the effects of leucines, non-metabolizable leucine analogues and other insulin secretagogues on the activity of glutamate dehydrogenase.

Glutamate dehydrogenase (GLDH) from bovine liver was employed in model system for testing a possible role of GLDH in insulin release. The ability of different insulin secretagogues to stimulate the activity of the diethylstilbestrol-inhibited enzyme was tested. The two insulin-releasing amino acids, L-leucine and its non-metabolizable analogue 2-aminobicyclo(2, 2, 1)heptane-2-carboxylic acid [b(--)-BCH], were the best stimulators of GLDH activity. The non-secreting stereoisomers, D-leucine and b(+)-BCH, were less effective. Glucose, L-arginine and the leucine metabolite alpha-ketoisocaproic acid lacked significant effects on GLDH activity. Small and diverging effects were obtained with sulfonvlurea compounds: whereas carbutamide caused slight stimulation, tolbutamide and glipizide had no effect, and glibenclamide was an inhibitor. The specificity of the insulin-releasing amino acids L-leucine and b(--)-BCH in stimulating GLDH activity makes it tempting to speculate about a connection between allosteric regulation of pyridine nucleotide-dependent enzymes and insulin release.

Amino Acids↗
PubMed2000

Epilithic and planktonic leucine aminopeptidase activity and leucine assimilation along the River Tweed, Scottish Borders.

The microbial organic-nitrogen transformation variables, extracellular leucine aminopeptidase activity, and leucine assimilation, were determined at stone surfaces and in river water at six sites along 138 km of the River Tweed and also in a major tributary, the River Teviot. Sampling was on 3 days, representative of spring, summer and autumn. The variables, except for epilithic leucine assimilation, all showed significant downstream increase on at least 2 of the sampling days. Their values tended to be lowest in spring (early April). Aminopeptidase activity, leucine assimilation, microbial abundance, and pattern of downstream increase in the Tweed were shown to be broadly similar to in the River Swale, northern England. Microbial organic-nitrogen transformations and abundance in the Teviot tended to equal or exceed those in the lower Tweed, probably because of enrichment of the tributary by sewage-works discharges.

Animals↗
PubMed1995

Two-metal ion mechanism of bovine lens leucine aminopeptidase: active site solvent structure and binding mode of L-leucinal, a gem-diolate transition state analogue, by X-ray crystallography.

The three-dimensional structures of bovine lens leucine aminopeptidase (blLAP) complexed with L-leucinal and of the unliganded enzyme have been determined at crystallographic resolutions of 1.9 and 1.6 A, respectively. Leucinal binds as a hydrated gem-diol to the active site of b1LAP), resembling the presumed gem-diolated intermediate in the catalytic pathway. One hydroxyl group bridges the two active site metal ions, and the other OH group is coordinated to Zn1. The high-resolution structure of the unliganded enzyme reveals one metal-bound water ligand, which is bridging both zinc ions. Together, these structures support a mechanism in which the bridging water ligand is the attacking hydroxide ion nucleophile. The gem-diolate intermediate is probably stabilized by four coordinating bonds to the dizinc center and by interaction with Lys-262 and Arg-336. In the mechanism, Lys-262 polarizes the peptide carbonyl group, which is also coordinated to Zn1. The Arg-336 side chain interacts with the substrate and the gem-diolate intermediate via water molecules. Near Arg-336 in the b1LAP-leucinal structure, an unusually short hydrogen bond is found between two active site water molecules.

Animals↗
PubMed1981

In vitro conversion of leucine to valine: configurational assignment of [5-13C]leucines.

Chiral (2S)-[5-13C]leucine was obtained from Escherichia coli deficient in the synthesis of acetolactate when cultures were supplemented with (RS)-[2-13CH3]acetolactate. The carbon-13 nuclear magnetic resonance spectrum showed one strong peak with a chemical shift of 21.4 ppm relative to tetramethylsilane [Sylvester, S. R., & Stevens, C. M. (1979) Biochemistry 18, 4529-4531]. Silver picolinate oxidation of the labeled leucine gave isovaleric acid which was then brominated at the alpha position to give (2RS)-2-bromo[3-13CH3]-isovaleric acid (2-bromo-3-[13C]methylbutanoic acid). Aminolysis afforded (2RS)-[4-13C]valine which was treated with D-amino acid oxidase in the presence of catalase. The final product was identified as (2S,3S)-[4-13C]valine by the specificity of D-amino acid oxidase, by amino acid analysis, and by the persistence of a strong signal at gamma 17.8 in the carbon-13 magnetic resonance spectrum. These results establish the absolute configuration of the biosynthetic leucine to be (2S,4S)-[5-13C]leucine.

Carbon Isotopes↗
PubMed1996

The carboxyl terminus of GLUT4 contains a serine-leucine-leucine sequence that functions as a potent internalization motif in Chinese hamster ovary cells.

To characterize the trafficking motifs contained in the carboxyl terminus of GLUT4, a chimera (GTCTR) was constructed in which the carboxyl-terminal 30 amino acids of GLUT4 were substituted for the amino-terminal cytoplasmic domain of the transferrin receptor (TR). The endocytic behavior of this chimera was characterized in Chinese hamster ovary cells. The GTCTR chimera had a more predominant intracellular distribution compared to the TR. Only 20% of the GTCTR chimera is on the surface at steady-state compared to 35% of the TR. The GTCTR chimera is internalized 50% more rapidly and recycled 20% more slowly than the TR. Acidification of the cytosol inhibited internalization of the GTCTR chimera, indicating that the chimera is internalized through clathrin-coated pits. Mutations of GTCTR were constructed in which a di-leucine sequence of the carboxyl domain of GLUT4 was mutated to a di-alanine sequence (GTCTR-AA) and serine residue 488, immediately preceding the di-leucine sequence, was mutated to either an alanine or aspartate residue. In each case, albeit to varying degrees, the substitutions shifted the distribution of the mutated GTCTR constructs toward the surface. The shift in the distribution of GTCTR-AA resulted from a 10-fold reduction in internalization, and the shift of serine 488 mutants resulted from a 3-fold reduction in the internalization rate compared to GTCTR. None of these mutations affected the recycling rate. These results demonstrate that the carboxyl terminus of GLUT4 contains a serine-leucine-leucine-based motif that, when expressed in non-insulin responsive cells, functions as a potent internalization motif which promotes more rapid internalization than does the native TR internalization motif.

Amino Acid Sequence↗
PubMed1978

Effect of leucine at different levels of pyridoxine on hepatic quinolinate phosphoribosyl transferase (EC 2.4.2.19) and leucine aminotransferase (EC 2.6.1.6) in rats.

1. Effects of incorporating 30 g leucine/kg into diets on quinolinate phosphoribosyl transferase (QPRT; EC 2.4.2.19) activity and leucine aminotransferase (EC 2.6.1.6) activity were studied in groups of rats receiving 5, 30 and 60 micrograms of pyridoxine/10 g diet. 2. The results indicated that 30 g leucine/kg diet significantly reduced the QPRT activity when the diets provided 5 micrograms pyridoxine/10 g and that the effect was only marginal when the diet included 30 micrograms pyridoxine/10 g. The inhibitory effect was completely absent when the diet provided higher amounts of pyridoxine (60 microgram/10 g). 3. These results suggest that additional amounts of pyridoxine are necessary to counteract the effects of excess of leucine in the diet. 4. Leucine aminotransferase activity was increased in rats given diets containing higher amounts of pyridoxine; supplementary leucine also increased the enzyme activity.

Animals↗
PubMed1989

Effects of a dietary excess of leucine and of the addition of leucine and 2-oxo-isocaproate on the metabolism of tryptophan and niacin in isolated rat liver cells.

1. Feeding rats on a low-tryptophan, niacin-free, high-leucine diet resulted in impaired synthesis from tryptophan of the nicotinamide nucleotide coenzymes, NAD and NADP, and N1-methyl nicotinamide in isolated hepatocytes, compared with cells from animals fed on a low-tryptophan, niacin-free control diet providing an appropriate amount of leucine. This was accompanied by reduced accumulation of the tryptophan metabolites kynurenine, 3-hydroxykynurenine and xanthurenic acid. 2. With hepatocytes from animals fed on the low-tryptophan, niacin-free control diet, the addition of leucine to the incubation medium resulted in reduced synthesis of niacin from tryptophan, and a small increase in the accumulation of 3-hydroxykynurenine. 3. With hepatocytes from animals fed on the low-tryptophan, niacin-free control diet, the addition of 2-oxoisocaproate to the incubation medium resulted in increased synthesis of NAD(P) and niacin, and increased accumulation of 3-hydroxykynurenine. 4. The results suggest that a dietary excess of leucine alters the activity of the enzymes of tryptophan----niacin metabolism, or the uptake of tryptophan into the liver, in a different manner from the simple inhibition and activation seen in experiments in vitro. 5. Differences between studies in isolated hepatocytes and intact animals suggest that extra-hepatic metabolism of tryptophan, catalysed by indoleamine dioxygenase, is more important than has been believed hitherto.

Animals↗
PubMed1977

[Secretion of radioactively labeled amino acids into the digestive tract. 2. 14C labeling and amount of 14C leucine in the intestinal contents after subcutaneous injection of 14C leucine in rats].

Experimental rats received a subcutaneous injection of 14C of varying activity. The rats were then killed in groups within a period of from 2 mins to 30 mins after the injection. A certain amount of 14C activity was detected in the intestinal contents as early as 2 mins after the injection, both as free 14C leucine and in the TCE soluble portion of the intestinal contents (TCE=trichloroacetic acid). A comparison of the degree of labelling in the TCE soluble and the TCE precipitable fractions of the intestinal contents showed that the most likely way for free 14C leucine to get into the lumen of the intestine is via the pancreatic juice while that for protein-bound 14C leucine is through the proteins in the pancreatic gland. The degree of labelling in both fractions decreased very steeply from the first to the third third of the intestine. This may be caused either by absorption of the secreted 14C leucine or may be brought about by a delay in food passage through the intestine.

Amino Acids↗
PubMed1981

[Absorption of 14C-leucine and 15N-leucine in rats after feeding a fish meal diet in comparison with a gelatin diet].

Albino rats received after nine days of adaptation to a fish meal diet in comparison with a gelatin diet 14C-U-Leucine and 15N-L-leucine via a pellet made from the specific diet after food deprivation for 15 h. Thereafter, the experimental animals consumed the non-labelled experimental diet ad libitum. Five times after the pellet intake, 30 min, and 1, 2, 4 and 6 h after injection of the labelled food, four experimental rats were sacrificed. The contents of the digestive tract and tissue samples were examined for 14C and 15N. We detected to following percentages of 14C-intake or the intake of 15N-excess (15N') in the TCA-soluble fraction of the gastric contents: [Formula: see text] We found in the TCA-soluble fraction of digesta of small intestine the following percentage of intake: [Formula see text] Where these values regarded as non-absorbed leucine, the 14C values obtained up to four hour period of experiment would be too high. Presumably, they are in the case of both diets simulated by other 14C-metabolites which originate from the leucine catabolism and reach the intestinal lumen. Amino acids labelled with 15N should be preferred in studies on the absorption of amino acids because, in case of catabolism, the 15N-aminogroup is excreted mainly in the form of urea via urine.

Animal Feed↗
PubMed1986

Studies on the secretion of amino acids and of urea into the gastro intestinal tract of pigs. 1. Secretion of leucine into the stomach and upper part of the duodenum during continuous intravenous infusion of 14C-leucine.

Three pigs of 34 kg live weight were fitted with a re-entrant cannula in the duodenum, and with two catheters placed in the jugular vein and carotid artery. They were fed 1.2 kg/d of wheat-dried-skimmed milk diet. Digesta from the proximal duodenal cannula were collected for 12 h on 2 consecutive days; 50% were reintroduced into the respective distal cannula and 50% were stored at -20 degrees C. Three days later 14C-leucine was infused into the jugular vein for 12 h, starting with the morning meal. During this period the digesta from the proximal cannula were collected and stored for analysis while the digesta collected previously were reintroduced into the distal cannula. Blood samples were taken from the carotid artery. The total flow of duodenal digesta in 12 h was 5760 +/- 530 g. On average 70 percent of the radioactivity in digesta was associated with the TCA-precipitable fraction. During hours 0-3 and 11-12 of infusion 60-70% and 96-98% of the radioactivity in the TCA precipitable fractions was in leucine. In the TCA soluble fraction only 30-40 per cent of the radioactivity was associated with leucine. At the plateau 2.1 and 3.3% of infused 14C-leucine and of radioactivity were recovered in the duodenal digesta. The calculated amount of endogenous protein passing the duodenum was 20.4 g/d/pig. The results are discussed in relation to previous studies on protein synthesis and secretion in which 14C- and 15N-amino acids were used.

Amino Acids↗
PubMed2004

Heptaaquatetra-mu3-hydroxy-hexa-mu2-L-leucine-(L-leucine)tetraytterbium(III) tetrachlorozinc(II) trichlorohydroxyzinc(II) tetrachloride octahydrate.

The title bimetallic compound, [Yb4(mu3-OH)4(C6H13NO2)7(H2O)7][ZnCl4][ZnCl3(OH)]Cl4.8H2O, was synthesized at near physiological pH (6.0). The compound exhibits some novel structural features, including an asymmetric [Yb4(mu3-OH)4(L-leucine)7(H2O)7]8+ complex cation in which four OH groups act as bridging ligands, linking four Yb3+ cations into a Yb4O4 structural unit. Each pair of adjacent Yb3+ ions is further bridged by one carboxy group from a leucine ligand. Water molecules and a monodentate leucine ligand also coordinate to Yb3+ ions, completing their eight-coordinate square-antiprismatic coordination. The Yb4(mu3-OH)4(L-leucine)7(H2O)7]8+ cation, the [ZnCl4]2-, [ZnCl3OH]2- and Cl- anions, and the lattice water molecules are linked via hydrogen bonds.

Crystallography, X-Ray↗
PubMed1990

The leucine regulon of Escherichia coli K-12: a mutation in rblA alters expression of L-leucine-dependent metabolic operons.

We have isolated and characterized a highly pleiotropic Escherichia coli mutant affected in the activity of a number of enzymes involved in different metabolic pathways, all of which are regulated by leucine. Selected for its ability to grow with L-serine as sole carbon source, the rbl-1::Tn10 mutant had high levels of L-serine deaminase activity (due to increased transcription of the structural gene) and of another amino acid-degrading enzyme, L-threonine dehydrogenase, and decreased transcription of the operons serA and ilvIH, coding for biosynthetic enzymes. The rbl mutation suppressed the slow growth of a metK mutant, deficient in S-adenosylmethionine synthetase. Furthermore, metK mutants spontaneously accumulated faster-growing rbl-like derivatives, and a commonly used metK strain, RG62, carries such a mutation. The rbl gene is located near 20 min on the E. coli genetic map. All phenotypes of the rbl mutant could be observed in rbl+ strains cultivated in the presence of L-leucine, and exogenous L-leucine had little further effect on the rbl strains. We propose that the rbl gene product is the regulator of a global response to leucine.

Alcohol Oxidoreductases↗