Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: DrugBank:EXPT02079 (lysine)
 58,762 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The pyruvate dehydrogenase multienzyme complex from bovine kidney and heart is inactivated by treatment with pyridoxal 5'-phosphate and sodium cyanide or sodium borohydride. The site of this inhibition is the pyruvate dehydrogenase (E1) component of the complex. Inactivation of E1 by the pyridoxal phosphate-cyanide treatment was prevented by thiamin pyrophosphate. Equilibrium binding studies showed that E1 contains two thiamin pyrophosphate binding sites per molecule (alpha 2 beta 2) and that modification of E1 increased the dissociation constant (Kd) for thiamin pyrophosphate about 5-fold. Incorporation of approximately 2.4 equiv of 14CN per mole of E1 tetramer in the presence of pyridoxal phosphate resulted in about a 90% loss of E1 activity. Radioactivity was incorporated predominantly into the E1 alpha subunit. Radioactive N6-pyridoxyllysine was identified in an acid hydrolysate of the E1-pyridoxal phosphate complex that had been reduced with NaB3H4. The data are interpreted to indicate that in the presence of sodium cyanide or sodium borohydride, pyridoxal phosphate reacts with a lysine residue at or near the thiamin pyrophosphate binding site of E1. This binding site is apparently located on the alpha subunit.
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PMID:Active-site modification of mammalian pyruvate dehydrogenase by pyridoxal 5'-phosphate. 408 75

Cross-reacting beta(2) subunits (CRMs) were purified from eight trpB missense mutants to test for complementation in vitro after urea dissociation and reaggregation. One CRM (B290, demonstrating "repairability," i.e., the appearance of enzymatic activity on combination with alpha subunits) was clearly positive with four others, all "non-repairable" CRMs resulting from mutations at three different but neighboring sites. One complementing pair, B290-B248, was studied in more detail and found, upon mixing purified proteins, to give complementation in the absence of denaturants. Complementation activity was low in each case. To study the mechanism of the modest increases in activity, we used a reduced beta(2) subunit as an artificial CRM to form hybrids where both the amount of activity due to complementation and the amount of hybrid could be measured. (In a reduced beta(2) subunit, the two pyridoxal phosphate cofactors have been chemically reduced by sodium borohydride and are covalently attached to lysine residues. This abolishes activity in the tryptophan synthetic reaction and causes the protein to migrate much faster than normal in acrylamide gel electrophoresis.) Reduced beta(2) subunit formed hybrid dimers with the non-repairable CRMs B244 and B248 at pH 6.0, but no enzymatic activity appeared. On the other hand, when reduced beta(2) subunit was mixed with B290 CRM at pH 6.0 to 6.6, an activity increase was seen that was proportional to the amount of hybrid. We conclude that hybrid formation is essential for complementation and that the mechanism of complementation in this system is the correction of a repairable active site on the B290 beta chain by a conformational change occuring when hybrid dimer is formed. This type of complementation must be restricted to a small class of CRMs having a conformationally deformed active site. From the amount of hybrid present and the increase in activity, a specific activity of 50 U/mg was calculated for the hybrid containing reduced and B290 beta chains. This value is slightly less than but close to the activity of the hybrid formed between reduced and normal beta chains, shown earlier to have half the specific activity of the normal dimer.
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PMID:Complementation in vitro between mutationally altered beta2 subunits of Escherichia coli tryptophan synthetase. 459 48

1. Bison ribonuclease was isolated from pancreas glands of Bison bison by acid extraction, (NH(4))(2)SO(4) fractionation, affinity chromatography on Sepharose-5'-(4-aminophenylphosphoryl)uridine 2',3'-phosphate and ion-exchange chromatography on Bio-Rex-70. 2. The selectivity of the affinity column towards bison ribonuclease in heterogeneous protein solutions was greatly improved by employing piperazine buffers at pH5.3, which decreased non-specific interactions of other proteins. Rapid desorption from the affinity column was obtained with sodium phosphate buffer (pH3). 3. Bison ribonuclease has a total amino acid content very similar to ox ribonuclease. Inactivation of bison ribonuclease with iodoacetic acid leads to the formation of 0.62 residues of pi-carboxymethylhistidine and 0.36 residues of tau-carboxymethylhistidine. The amino acid composition of peptides isolated from diagonal peptide ;maps' and also of peptides isolated after pH1.6 and 2.4 two-dimensional high-voltage electrophoresis of a digest of bison ribonuclease labelled with pyridoxal 5-phosphate indicates that there is complete homology between ox and bison ribonucleases. 4. The Schiff-base attachment site of pyridoxal 5-phosphate was identified as lysine-41 by NaBH(4) reduction followed by peptide isolation.
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PMID:The isolation and partial characterization of ribonuclease A from Bison bison. 477 70

Two distinct phenotypic classes of lysine requiring auxotrophs of Escherichia coli are described. Mutants of the LysA class produce little or no active diaminopimelic acid (DAP) decarboxylase and specifically require lysine for growth. Mutants of the LysB class produce a cryptic DAP decarboxylase which can be activated both in vivo and in vitro by higher than normal levels of its cofactor, pyridoxal 5'-phosphate. The LysB mutants have an alternate requirement for lysine or pyridoxine. Both LysA and LysB mutations map at 55 min, close to the thyA locus of E. coli. The association between pyridoxal phosphate and DAP decarboxylase appears to be much weaker in LysB mutants than in wild-type bacteria, and the mutant enzyme also sediments more slowly than wild-type enzyme in sucrose density gradients. The results suggest that the LysB mutations alter a specific region (or subunit) of the enzyme molecule which is needed to stabilize the binding of pyridoxal phosphate. These studies help to resolve certain contradictory observations on DAP decarboxylase reported earlier and may have relevance to pyridoxal phosphate enzymes in general. Prototrophic revertants of LysB mutants arise by second site mutations that result in increased availability of intracellular pyridoxal phosphate. These revertants appear to be derepressed for pyridoxine biosynthesis.
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PMID:Mutants of Escherichia coli with a growth requirement for either lysine or pyridoxine. 492 90

An Escherichia coli mutant resistant to isoniazid (WG497) contained 0.6 mumole of extracellular pyridoxamine and pyridoxamine phosphate in the early stationary phase. A suppressed lysine mutant (AT1024) contained 1.4 mumoles of pyridoxal phosphate under the same conditions. The internal concentration of vitamin B(6) was one-half of normal for AT1024 and increased fivefold for WG497.
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PMID:Identification of the forms of vitamin B 6 present in the culture media of "vitamin B 6 control" mutants. 493 69

The meso-diaminopimelate (DAP) decarboxylase of Bacillus licheniformis, a pyridoxal phosphate-requiring enzyme, was stabilized in vitro by 0.15 m sodium phosphate buffer (pH 7.0) containing 1 mm 2,3-dimercaptopropan-1-ol, 100 mug of pyridoxal phosphate per ml, and 3 mm DAP. When the meso-DAP concentration was varied, the enzyme in cell-free extracts of B. licheniformis exhibited Michaelis-Menten kinetics. Pyridoxal phosphate was the only pyridoxine derivative which acted as a cofactor. The enzyme was subject to both inhibition and repression by l-lysine. The inhibitory effect of lysine was on the K(m) (meso-DAP). A maximum repression of about 20% was obtained. No significant inhibition or activation was produced by cadaverine, dipicolinic acid, phenylalanine, pyruvate, ethylenediamine-tetraacetate, adenosine triphosphate, adenosine diphosphate, or adenosine monophosphate. When B. licheniformis was grown in an ammonium lactate-glucose-salts medium, an increase in DAP decarboxylase specific activity occurred during cellular growth with a maximal specific activity at the end of the exponential phase. As soon as growth ceased, the specific activity of the enzyme decreased to approximately one-half of the maximal specific activity and remained at this level thereafter. When B. cereus was grown in complex media, there was an increase in DAP decarboxylase specific activity up to the end of the exponential phase. Thereafter, the specific activity decreased to a nondetectable level in 4 hr. Dipicolinic acid synthesis was first detected 15 min later and was essentially complete after an additional 2.5 hr. The significance of the disappearance of DAP decarboxylase in B. cereus was discussed with regard to control of dipicolinic acid and spore mucopeptide biosynthesis.
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PMID:Diaminopimelate decarboxylase of sporulating bacteria. 497 17

1. Acetylation of aspartate aminotransferase from pig heart inhibits completely the enzymic activity when the coenzyme is in the amino form (pyridoxamine phosphate) or when the coenzyme has been removed, but not when the coenzyme is in the aldehyde form (pyridoxal phosphate). 2. The group the acylation of which is responsible for the inhibition has been identified with the in-amino group of a lysine residue at the coenzyme-binding site. Moreover, in the pyridoxamine-enzyme the amino group of the coenzyme is also acetylated. 3. The reactivity of the coenzyme-binding lysine residue is greatly different in the pyridoxamine-enzyme and in the apoenzyme, suggesting the possibility of an interaction of its in-amino group with pyridoxamine or with other groups on the protein.
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PMID:Acylation of aspartate aminotransferase. 604 35

An increase in pH shifts the equilibrium between the K+-form and the Na+-form of the (Na+ + K+)-ATPase towards the Na+-form. pK for the proton effect on the equilibrium is decreased by modification of the enzyme with pyridoxal 5-phosphate. The reactivity of the enzyme towards pyridoxal 5-phosphate is increased by an increase in pH. Modification by pyridoxal 5-phosphate of epsilon-amino groups on lysine, which has a pK of about 8 with the enzyme in the K+-form and of about 7.4 in the Na+-form, shifts the equilibrium between E1Na+ and E2 towards E2, and the equilibrium between E2(K+occ) and E2 towards E2, but has no effect on the overall equilibrium between E1Na+ and E2(K+occ). An additional modification of epsilon-amino groups on lysine, which has a pK of 9.5-10 with the enzyme in the K+-form and of about 7.7 with the enzyme in the Na+-form, shifts the equilibrium between E2(K+occ) and E1Na+ towards E1Na+; this is due to a shift in the equilibrium between E2(K+occ) and E2 towards E2, but with no effect on the equilibrium between E1Na+ and E2. The results show that the transition from the K+-form to the Na+-form decreases the pK of lysine epsilon-amino groups on the enzyme, and that the protonation of these groups influences the equilibrium between the two conformations.
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PMID:Effect on the equilibrium between the Na+-form and the K+-form of the (Na+ + K+)-ATPase of modification of the enzyme with pyridoxal 5-phosphate. 608 13

The pyridoxal phosphate (PLP) modification of the lysine amino groups in cytochrome c causes decrease in the reaction rate with cytochrome c oxidase. The rate constants for (PLP)2-cyt. c, PLP(Lys 86)-cyt. c, PLP(Lys 79)-cyt. c and native cytochrome c (at pH 7.4, I = 0.02) are 3.6 X 10(-3) sec-1, 5.5 X 10(-3) sec-1, 5.2 X 10(-3) sec-1 and 9.8 X 10(-3) sec-1, respectively. In spite of the same positive charge of singly PLP-cytochromes c the reaction between PLP(Lys 86)-cyt. c and cyt. c oxidase exhibits the ionic strength dependence that differs from those of the PLP(Lys 79)-cyt. c. The rate constants at zero and infinite ionic strength for PLP(Lys 86)-cyt. c is 2-fold less than that for PLP(Lys 79)-cyt. c. The positively charged cytochrome c lysines 86 and 79 form two from four or five predicted complementary charge interactions with carboxyl groups on cytochrome c oxidase.
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PMID:Charge interactions of cytochrome c with cytochrome c oxidase. 609 87

Modification of yeast phosphofructokinase (E.C. 2.7.1.11) with pyridoxal 5'-phosphate leads to a decreased enzyme activity. ATP at higher concentrations protects the enzyme against inactivation, while fructose 6-phosphate has no effect. At relatively low concentrations of pyridoxal phosphate the inhibition of activity by ATP was decreased or even abolished. AMP still activates the modified enzyme pointing to a separate binding site in the regulatory centre. The dissociation of the phosphopyridoxyl enzyme in products with apparent sedimentation coefficients of about 4-6 S gives evidence that lysine might be involved in the stabilization of the quaternary structure of yeast phosphofructokinase. The incorporation of pyridoxal phosphate in the 17 S form of yeast phosphofructokinase as obtained by a partial proteolysis does not show differences from the 20 S, proteolytically unmodified enzyme.
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PMID:Modification of yeast phosphofructokinase with pyridoxal 5'-phosphate. 623 Oct 21


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