Kinetics of the inactivation of Escherichia coli glutamate apodecarboxylase by phenylglyoxal

Cheung, Shu-Tong; Fonda, Margaret L.

doi:10.1016/0003-9861(79)90529-0

Cited by 18 publications

(6 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…6 and 9) . This selectivity can be compared to the remarkable selectivity of phenylglyoxal for modification of only the functionally essential arginyl residues at enzyme active sites, as many enzymes can be fully inactivated by phenylglyoxal after modification of a single arginyl residue per enzyme protomer or subunit (Borders and Riordan, 1975 ;Berghauser, 1975 ;Armstrong et al, 1976 ;Kantrowitz and Lipscomb, 1976 ;Philips et al, 1979;Cheung and Fonda, 19796;Borders and Johansen, 1980a, b ;Vandenbunder et al, 1981) . Phenylglyoxal is not an "affinity" reagent or an "active-site-directed" reagent, for most studies using this reagent show that rates of inactivation of enzymes are first order in phenylglyoxal.…”

Section: Stoichiometry Between Phenylglyoxal Binding and Transport Inactivationmentioning

confidence: 99%

Selective phenylglyoxalation of functionally essential arginyl residues in the erythrocyte anion transport protein.

Bjerrum¹,

Wieth²,

Borders³

1983

The Journal of General Physiology

View full text Add to dashboard Cite

The red cell anion transport protein, band 3, can be selectively modified with phenylglyoxal, which modifies arginyl residues (arg) in proteins, usually with a phenylglyoxal :arg stoichiometry of 2 :1 . Indiscriminate modification of all arg in red cell membrane proteins occurred rapidly when both extra-and intracellular pH were above 10 . Selective modification of extracellularly exposed arg was achieved when ghosts with a neutral or acid intracellular pH were treated with phenylglyoxal in an alkaline medium . The rate and specificity of modification depend on the extracellular chloride concentration . At 165 mM chloride maximum transport inactivation was accompanied by the binding of four phenylglyoxals per band 3 molecule. After removal of extracellular chloride, maximum transport inhibition was accompanied by the incorporation of two phenylglyoxals per band 3, which suggests that transport function is inactivated by the modification of a single arg . After cleavage of band 3 with extracellular chymotrypsin, [14 C]phenylglyoxal was located almost exclusively in a 35,000-dalton peptide . In contrast, the primary covalent binding site of the isothiocyanostilbenedisulfonates is a lysyl residue in the second cleavage product, a 65,000-dalton fragment. This finding supports the view that the transport region of band 3 is composed of strands from both chymotryptic fragments . The binding of phenylglyoxal and the stilbene inhibitors interfered with each other. The rate of phenylglyoxal binding was reduced by a reversibly binding stilbenedisulfonate (DNDS), and covalent binding of [3 H]DIDS to phenylglyoxal-modified membranes was strongly delayed . At DIDS concentrations below 10 j,M, only 50% of the band 3 molecules were labeled with [ 3H]-DIDS during 90 min at 38°C, thereby demonstrating an interaction between binding of the two inhibitors to the protomers of the oligomeric band 3 molecules .

show abstract

Section: Stoichiometry Between Phenylglyoxal Binding and Transport Inactivationmentioning

confidence: 99%

Selective phenylglyoxalation of functionally essential arginyl residues in the erythrocyte anion transport protein.

Bjerrum¹,

Wieth²,

Borders³

1983

The Journal of General Physiology

View full text Add to dashboard Cite

show abstract

“…A number of pyridoxal phosphate enzymes have also been studied with dicarbonyl reagents. In some of these enzymes, the phosphate of the coenzyme binds to a specific arginyl residue; these enzymes include -aminobutyrate aminotransferase (Tunnicliff, 1980), glutamate decarboxylase (Cheung & Fonda, 1979), and o-serine dehydratase (Kazarinoff & Snell, 1976).…”

mentioning

confidence: 99%

L-Serine binds to arginine-148 of the .beta.2 subunit of Escherichia coli tryptophan synthase

Tanizawa¹,

Miles²

1983

Biochemistry

View full text Add to dashboard Cite

Inactivation of the beta 2 subunit and of the alpha 2 beta 2 complex of tryptophan synthase of Escherichia coli by the arginine-specific dicarbonyl reagent phenylglyoxal results from modification of one arginyl residue per beta monomer. The substrate L-serine protects the holo beta 2 subunit and the holo alpha 2 beta 2 complex from both inactivation and arginine modification but has no effect on the inactivation or modification of the apo forms of the enzyme. This result and the finding that phenylglyoxal competes with L-serine in reactions catalyzed by both the holo beta 2 subunit and the holo alpha 2 beta 2 complex indicate that L-serine and phenylglyoxal both bind to the same essential arginyl residue in the holo beta 2 subunit. The apo beta 2 subunit is protected from phenylglyoxal inactivation much more effectively by phosphopyridoxyl-L-serine than by either pyridoxal phosphate or pyridoxine phosphate, both of which lack the L-serine moiety. The phenylglyoxal-modified apo beta 2 subunit binds pyridoxal phosphate and the alpha subunit but cannot bind L-serine or L-tryptophan. We conclude that the alpha-carboxyl group of L-serine and not the phosphate of pyridoxal phosphate binds to the essential arginyl residue in the beta 2 subunit. The specific arginyl residue in the beta 2 subunit which is protected by L-serine from modification by phenyl[2-14C]glyoxal has been identified as arginine-148 by isolating a labeled cyanogen bromide fragment (residues 135-149) and by digesting this fragment with pepsin to yield the labeled dipeptide arginine-methionine (residues 148-149). The primary sequence near arginine-148 contains three other basic residues (lysine-137, arginine-141, and arginine-150) which may facilitate anion binding and increase the reactivity of arginine-148. The conservation of the arginine residues 141, 148, and 150 in the sequences of tryptophan synthase from E. coli, Salmonella typhimurium, and yeast supports a functional role for these three residues in anion binding. The location and role of the active-site arginyl residues in the beta 2 subunit and in two other enzymes which contain pyridoxal phosphate, aspartate aminotransferase and glycogen phosphorylase, are compared.

show abstract

“…Arg-406 is involved in binding the L-isovaline carboxylate group in dialkylglycine decarboxylase (Toney et al, 1993), while Arg-292 and Arg-386 interact with the substrate at the co-and a-carboxyl group, respectively, in aspartate aminotransferase (Almo et al, 1994). In addition, evidence from chemical modification and site-directed mutagenesis studies has implicated the involvement of arginine residues in the active site of several PLP-dependent enzymes (Kazarinoff and Snell, 1977;Cheung and Fonda, 1979;Tunnicliff, 1980;Schnackerz and Snell, 1983;Tanizawa and Miles, 1983;Marceau et al, 1989). In E. coli D-serine dehydratase, a single arginine residue near the active site was shown to be important for both the cofactor affinity and catalytic activity;…”

Section: Discussionmentioning

confidence: 99%

Aminolevulinate synthase: functionally important residues at a glycine loop, a putative pyridoxal phosphate cofactor-binding site

Gong

Ferreira²

1995

Biochemistry

View full text Add to dashboard Cite

5-Aminolevulinate synthase catalyzes the first step of the heme biosynthetic pathway in nonplant higher eukaryotes. The enzyme functions as a homodimer and requires pyridoxal 5'-phosphate as its cofactor. Lysine-313 in murine erythroid aminolevulinate synthase has been identified as the residue involved in the Schiff base linkage with pyridoxal 5'-phosphate [Ferreira, G. C., Neame, P. J., & Dailey, H. A. (1993) Protein Sci. 2, 1959-1965]. However, other residues involved in binding and orienting the cofactor remain unknown. We studied the informational content of each residue within an 11 amino acid glycine-rich region, which we propose to be part of the phosphate-binding motif, based on amino acid sequence comparison with other pyridoxal 5'-phosphate-dependent enzymes and nucleotide-binding proteins. Partial random mutagenesis of this region in murine erythroid aminolevulinate synthase gene was followed by an efficient biological selection, using a hemA- Escherichia coli strain to recover functional unnatural enzymes. Among the total of 5444 variants produced, 283 were found to be functional. DNA sequencing results of 226 functional mutants indicated that most residues in this region contained a low informational content, being able to tolerate several other amino acid substitutions. However, three residues, namely, Arg-149, Gly-142, and Gly-144, were found to contain high informational content; Arg-149 was conserved in all of the functional mutants sequenced, while Gly-142 and Gly-144 could only tolerate alanine replacement. Two codon-specific random libraries of Arg-149, and Gly-142 and -144, respectively, were constructed to test further the stringency of these three positions.(ABSTRACT TRUNCATED AT 250 WORDS)

show abstract

Kinetics of the inactivation of Escherichia coli glutamate apodecarboxylase by phenylglyoxal

Cited by 18 publications

References 43 publications

Selective phenylglyoxalation of functionally essential arginyl residues in the erythrocyte anion transport protein.

Selective phenylglyoxalation of functionally essential arginyl residues in the erythrocyte anion transport protein.

L-Serine binds to arginine-148 of the .beta.2 subunit of Escherichia coli tryptophan synthase

Aminolevulinate synthase: functionally important residues at a glycine loop, a putative pyridoxal phosphate cofactor-binding site

Contact Info

Product

Resources

About