Studies on adenosine triphosphate transphosphorylases XIV. Equilibrium binding properties of the crystalline rabbit and calf muscle ATP-AMP transphosphorylase (adenylate kinase) and derived peptide fragments

Hamada, Minoru; Palmieri, Richard H.; Russell, Geoffrey A.; Kuby, Stephen A.

doi:10.1016/0003-9861(79)90338-2

Cited by 62 publications

(63 citation statements)

References 35 publications

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“…The triphosphate moiety is located between the side chains of Lys-21, Gln-24, and Lys-27, and a hydrophobic sequence, residues 114-118, terminated by an anionic residue, Asp-119. In this orientation the y-phosphoryl group points toward the C-terminal 23-amino acid residues of the enzyme; a peptide corresponding to this region selectively binds 1,N6-ethenoadenosine monophosphate (EAMP) with substantial affinity (13).…”

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confidence: 99%

“…We have identified a third homologous region in several of these proteins and have further expanded the list of proteins with sequence homologies to adenylate kinase. Through a series of NMR studies on porcine (11) and rabbit muscle (12) High-field proton NMR was used to study the interaction of metal-ATP substrates with porcine (11) and rabbit muscle adenylate kinase (12), and with a globular peptide fragment of the latter enzyme consisting of residues 1-45 that binds metal-ATP with comparable affinity (13 (32).…”

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confidence: 99%

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ATP-binding site of adenylate kinase: mechanistic implications of its homology with ras-encoded p21, F1-ATPase, and other nucleotide-binding proteins.

Fry¹,

Kuby²,

Mildvan³

1986

Proc. Natl. Acad. Sci. U.S.A.

503

252

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The MgATP binding site of adenylate kinase, located by a combination of NMR and x-ray diffraction, is near three protein segments, five to seven amino acids in length, that are homologous in sequence to segments found in other nucleotide-binding phosphotransferases, such as myosin and Fl-ATPase, ras p21 and transducin GTPases, and cAMPdependent and src protein kinases, suggesting equivalent mechanistic roles of these segments in all of these proteins. Segment 1 is a glycine-rich flexible loop that, on adenylate kinase, may control access to the ATP-binding site by changing its conformation. Segment 2 is an a-helix containing two hydrophobic residues that interact with the adenine-ribose moiety of ATP, and a lysine that may bind to the j8-and y-phosphates of ATP. Segment 3 is a hydrophobic strand of parallel a6-pleated sheet, terminated by a carboxylate, that flanks the triphosphate binding site. The various reported mutations of ras p21 that convert it to a transforming agent all appear to involve segment 1, and such substitutions may alter the properties of p21 by hindering a conformational change at this segment. In F1-ATPase, the flexible loop may, by its position, control both the accessibility and the ATP/ADP equilibrium constant on the enzyme.Adenylate kinase catalyzes the reversible transfer of a phosphoryl group from MgATP to AMP.MgATP + AMP MgADP + ADP.[1]The enzyme has been purified from many sources, the x-ray structure of the porcine muscle enzyme has been reported at 3 A resolution (1), and amino acid sequences have been determined for muscle adenylate kinase from pig (2), human (3), calf, and rabbit (4). It was noted by Walker et al. (5) that two portions of the adenylate kinase sequence had homologous counterparts in the sequences of several ATPases. One of these homologous segments was later found to be present in p21, the GTPase that is the product of the ras oncogene (6-8). The recent finding that a mutation in this segment is the basis of the transforming ability of this protein (9, 10) has amplified the importance of determining the functional role of this segment. We have identified a third homologous region in several of these proteins and have further expanded the list of proteins with sequence homologies to adenylate kinase. Through a series of NMR studies on porcine (11) and rabbit muscle (12) High-field proton NMR was used to study the interaction of metal-ATP substrates with porcine (11) and rabbit muscle adenylate kinase (12), and with a globular peptide fragment of the latter enzyme consisting of residues 1-45 that binds metal-ATP with comparable affinity (13). Paramagnetic effects of B,y-bidentate Cr3+-ATP on the relaxation rates of protons of the enzyme and the peptide were measured and provided a total of eight distances from Cr3+ to the side chains of specific amino acid residues. Time-dependent nuclear tIn some of the proteins of (32). 907The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "adv...

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confidence: 99%

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confidence: 99%

ATP-binding site of adenylate kinase: mechanistic implications of its homology with ras-encoded p21, F1-ATPase, and other nucleotide-binding proteins.

Fry¹,

Kuby²,

Mildvan³

1986

Proc. Natl. Acad. Sci. U.S.A.

503

252

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“…There are two distinct nucleotide-binding sites in AK, one for MgATP 2-or MgADP 3-, and the other for AMP2-or ADP- (3,4). AK is often cited as a good model of a typical ATP-binding protein when structural topological comparisons or amino acid homology comparisons are made of the nucleotide-binding proteins (5).…”

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confidence: 99%

Site‐directed mutagenesis and steady‐state kinetic analysis of mutant enzymes of human adenylate kinase

et al. 1998

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SUMMARY:Site-directed mutagenesis of human adenylate kinase (AK) was carried out on residues His36, Lys55, and the C-terminal segment (Vail82, V186, and Leu193). Five mutants { (H36T, K55G, V 182G, V 186S, and L 193Stop (deletion of residues 193-194) } were generated and analyzed by steady-state kinetics. H36T, K55G, and L193Stop mutants showed an increase of Krn values (19.8-, 19.7-, and 11.3-fold) for AMP 2-compared to that for the wild-type enzyme, and these residues appeared to interact with AMP2-. V182G showed an increased Km value (7.4-fold) for MgATP 2-. Therefore, V182 may be essential for interaction with MgATP 2-. V186S increased the Km value (7.0-and 7.5-fold) for MgATP 2-and AMP 2-. V186 may thus interact with both substrates. The C-terminal domain of AK appears to be essential for MgATP 2-and AMP 2-binding.

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“…Hamada et al (35) have shown that a tryptic peptide containing the NH2-terminal 44 residues from rabbit muscle adenylate kinase binds specifically to the fluorescent ATP analog EATP (1,N6-etheno-ATP). Recently, Fry et al (36) have used NMR studies to demonstrate that the contacts between ATP and residues in this peptide are nearly identical to those between ATP and intact adenylate kinase and that these contacts can be accommodated by the crystallographic structure of the enzyme.…”

Section: Photoaffinity Labeling Of Tryptic Peptide T-31 Previouslymentioning

confidence: 99%

Nucleotide binding by a 24-residue peptide from the RecA protein of Escherichia coli.

Knight¹,

McEntee²

1986

Proc. Natl. Acad. Sci. U.S.A.

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We have recently demonstrated that two ATP analog affinity labels, 8-azidoadenosine 5'-triphosphate (N3ATP) and 5'-p-fluorosulfonylbenzoyladenosine (5'FSBA), covalently modify RecA protein ofEscherichia coli at a specific tyrosine residue (Tyr-264) located within a 24-residue tryptic peptide (T-31) spanning residues 257-280. Here we show that N3ATP efficiently modifies purified peptide T-31 and show that the interaction is specific by the following criteria: (i) photolabeling of peptide T-31 is saturable with respect to the N3ATP concentration; (ii) photolabeling is competitive with ATP and adenosine but not with adenine, UTP, or TTP; and (iii) other peptides derived from RecA protein were poor substrates for photolabeling except for one fragment that showed a nonspecific interaction with the photoaffinity analog. Analysis of N3ATP-modified T-31 shows that the photolabel attaches to more than one site within the peptide. These data argue that peptide T-31 contains some sites of contact for adenine and ribose moieties of ATP when it is bound to RecA protein.The RecA protein of Escherichia coli possesses DNAdependent nucleoside triphosphatase activity (1, 2). The binding and hydrolysis of certain nucleoside triphosphates are essential for other activities of this multifunctional enzyme. ATP greatly stimulates the ability of RecA protein to renature complementary single-stranded DNA chains (3,4) and is necessary for homologous pairing of single strands within duplex DNA molecules (5, 6). These reactions likely reflect the role of RecA protein in postreplication repair of damaged DNA and in homology-dependent recombination in cells (7,8). Moreover, RecA protein promotes the proteolytic inactivation of bacteriophage repressors (9) and a cellular repressor protein (LexA) that regulates expression of the "SOS" genes of E. coli (10). Both ATP (or the derivative These analogs were used to identify widely spaced residues in the 3-subunit of Fj-ATPase from bovine heart mitochondria (17, 18). That both of these affinity labels covalently modified the same tyrosine residue in the RecA protein suggested that residues contiguous with Tyr-264 could be sufficient for formation of a functional nucleotide binding site. We report here a series of experiments in which N3ATP was used to covalently modify purified peptide T-31. We demonstrate that N3ATP binding to T-31 is saturable and that ATP effectively competes for the site, suggesting that the binding of N3ATP to T-31 is specific. In addition, this specific interaction with N3ATP appears to be a unique property of T-31 when compared with the interaction ofN3ATP and other purified tryptic peptides derived from RecA (22). Photochemical crosslinking of N3ATP to purified tryptic peptides was routinely performed at room temperature in 50 mM Tris HCl (pH 7.5), 10 mM MgCl2, 20 mM KC1. Irradiation was done using an 8-W germicidal lamp 12 cm from the reaction mixtures. Measurements of covalent attachment of N3ATP to a given peptide were done using HPLC. At indicated times, aliq...

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Studies on adenosine triphosphate transphosphorylases XIV. Equilibrium binding properties of the crystalline rabbit and calf muscle ATP-AMP transphosphorylase (adenylate kinase) and derived peptide fragments

Cited by 62 publications

References 35 publications

ATP-binding site of adenylate kinase: mechanistic implications of its homology with ras-encoded p21, F1-ATPase, and other nucleotide-binding proteins.

ATP-binding site of adenylate kinase: mechanistic implications of its homology with ras-encoded p21, F1-ATPase, and other nucleotide-binding proteins.

Site‐directed mutagenesis and steady‐state kinetic analysis of mutant enzymes of human adenylate kinase

Nucleotide binding by a 24-residue peptide from the RecA protein of Escherichia coli.

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