Characterization and cAMP inhibition of a lysyl-(N-ϵ-5′-phospho) adenosyl phosphoamidase in Dictyostelium discoideum

Rossomando, Edward F.; Hadjimichael, Jane

doi:10.1016/0020-711x(86)90193-x

Cited by 13 publications

(8 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The supposition that NH 2 -pA occurs in other organisms and that its concentration can be enzymatically controlled is supported by the existence of adenosine-, or generally nucleoside-5′-phosphoramidate hydrolases (EC 3.9.1.-) that catalyze cleavage of the substrate P-N bond and liberate the corresponding 5′-NMP (pN) and ammonia from NH 2 -pA or other NH 2 -pNs. This catabolic activity has been demonstrated in D. discoideum (Rossomando & Hadjimichael, 1986), rat liver (Kuba et al, 1994), and appears to be a catalytic feature of various HIT-proteins (Bieganowski et al, 2002;Guranowski et al, 2008;2010a;2010b). Our preliminary studies showed that NH 2 -pA can be hydrolyzed by at least two enzymes from yellow lupin -dinucleoside triphosphatase/Fhit homolog protein (EC 3.6.1.29) and nucleoside phosphoramidase (Hint protein) (Guranowski et al, 2006).…”

Section: Introductionsupporting

confidence: 59%

Plant nucleoside 5'-phosphoramidate hydrolase; simple purification from yellow lupin (Lupinus luteus) seeds and properties of homogeneous enzyme.

et al. 2011

View full text Add to dashboard Cite

Adenosine 5'-phosphoramidate (NH₂-pA) is an uncommon natural nucleotide of poorly understood biochemistry and function. We studied a plant enzyme potentially involved in the catabolism of NH₂-pA. A fast and simple method comprising extraction of yellow lupin (Lupinus luteus) seed-meal with a low ionic strength buffer, ammonium sulfate and acetone fractionations, removal of contaminating proteins by heat denaturation, and affinity chromatography on AMP-agarose, yielded homogenous nucleoside 5'-phosphoramidase. Mass spectrometric analysis showed that the lupin hydrolase exhibits closest similarity to Arabidopsis thaliana Hint1 protein. The substrate specificity of the lupin enzyme, in particular its ability to split the P-S bond in adenosine 5'-phosphorothioate, is typical of known Hint1 proteins. Adenosine 5'-phosphofluoride and various derivatives of guanosine 5'-phosphoramidate were also substrates. Neither common divalent metal cations nor 10 mM EDTA or EGTA affected the hydrolysis of NH₂-pA. The enzyme functions as a homodimer (2 x 15,800 Da). At the optimum pH of 7.0, the K(m) for NH₂-pA was 0.5 µM and k(cat) 0.8 s⁻¹ (per monomer active site). The properties of the lupin nucleoside 5'-phosphoramidase are compared with those of its counterparts from other organisms.

show abstract

Section: Introductionsupporting

confidence: 59%

Plant nucleoside 5'-phosphoramidate hydrolase; simple purification from yellow lupin (Lupinus luteus) seeds and properties of homogeneous enzyme.

et al. 2011

View full text Add to dashboard Cite

show abstract

“…Enzymatic Assays-Compounds tested as potential substrates were from Sigma, except AMP-N-alanine methyl ester and AMP-N-⑀-(N-␣-acetyl lysine methyl ester), which were synthesized according to a modification of the synthesis of AMP-N-⑀-lysine (49), and human DNA ligase I adenylylated intermediate, a gift of Alan E. Tomkinson, which was prepared as described (50). Compounds were screened for hydrolysis in 10-l reactions containing 1 mM substrate, 0.5-2.0 g of enzyme, 0.5 mM MgCl 2 , 20 mM Na-PIPES at pH 7.2 for 5-12 h at 37°C.…”

Section: Purification Of Recombinant Hint and Hnt1mentioning

confidence: 99%

Adenosine Monophosphoramidase Activity of Hint and Hnt1 Supports Function of Kin28, Ccl1, and Tfb3

Bieganowski

Garrison

Hodawadekar

et al. 2002

Journal of Biological Chemistry

104

163

View full text Add to dashboard Cite

The histidine triad superfamily of nucleotide hydrolases and nucleotide transferases consists of a branch of proteins related to Hint and Aprataxin, a branch of Fhitrelated hydrolases, and a branch of galactose-1-phosphate uridylyltransferase (GalT)-related transferases. Although substrates of Fhit and GalT are known and consequences of mutations in Aprataxin, Fhit, and GalT are known, good substrates had not been reported for any member of the Hint branch, and mutational consequences were unknown for Hint orthologs, which are the most ancient and widespread proteins in the Hint branch and in the histidine triad superfamily. Here we show that rabbit and yeast Hint hydrolyze the natural product adenosine-5-monophosphoramidate (AMPNH 2 ) in an active-site-dependent manner at second order rates exceeding 1,000,000 M ؊1 s ؊1 . Yeast strains constructed with specific loss of the Hnt1 active site fail to grow on galactose at elevated temperatures. Loss of Hnt1 enzyme activity also leads to hypersensitivity to mutations in Ccl1, Tfb3, and Kin28, which constitute the TFIIK kinase subcomplex of general transcription factor TFIIH and to mutations in Cak1, which phosphorylates Kin28. The target of Hnt1 regulation in this pathway was shown to be downstream of Cak1 and not to affect stability of Kin28 monomers. Functional complementation of all Hnt1 phenotypes was provided by rabbit Hint, which is only 22% identical to yeast Hnt1 but has very similar adenosine monophosphoramidase activity. Histidine triad (HIT)1 proteins are a superfamily of nucleotide-binding proteins named for a near C-terminal HXHXHXX motif (X is a hydrophobic amino acid) positioned at the ␣-phosphate of nucleotide substrates (1). The first branch of the superfamily is named for rabbit Hint, which had been purified as an abundant protein from cardiac cytosol by adenosine affinity chromatography (2) and shown to have homologs in all forms of life (1). Recently, Aprataxin, a gene located at 9p13 that is inactivated in ataxia with oculomotor apraxia, the second most common of the autosomal recessive ataxias, was identified as a member of the Hint branch of the HIT superfamily (3, 4). Human Fhit (5), which functions as a tumor suppressor protein in human (6 -9) and murine (10, 11) epithelial tissues, is the prototypical member of the second branch of the HIT superfamily. Fhit homologs have been found in fungi (12) and animals (13-16) and exhibit diadenosine polyphosphate hydrolase activity. A third branch of the HIT superfamily contains more distantly related nucleotide transferases including galactose-1-phosphate uridylyltransferase, which is the enzyme deficient in galactosemics (1), budding yeast diadenosine tetraphosphate phosphorylases ApaI and Apa2 (17), and adenylylsulfate:phosphate adenylyltransferase (18). Ironically, although Hint is the most ancient and widespread of the HIT proteins, reasonable Hint substrates remained unidentified, and the consequences of mutations in Hint or Hint orthologs were unknown (19).Recently, human Hint was identified as...

show abstract

“…Traditionally compounds containing a nitrogen–phosphorus bond are divided into two separate classes: I and II [4,5]. Compounds of class I are characterized by the presence of terminal P–OH group (all N -phosphorylated amino acids in proteins belong to this group as well as many small molecules e.g., phosphagen— N -phosphocreatine).…”

Section: Natural Products Containing a P–n Bond (Phosphoramidates)mentioning

confidence: 99%

Natural Products Containing ‘Rare’ Organophosphorus Functional Groups

2019

View full text Add to dashboard Cite

Phosphorous-containing molecules are essential constituents of all living cells. While the phosphate functional group is very common in small molecule natural products, nucleic acids, and as chemical modification in protein and peptides, phosphorous can form P–N (phosphoramidate), P–S (phosphorothioate), and P–C (e.g., phosphonate and phosphinate) linkages. While rare, these moieties play critical roles in many processes and in all forms of life. In this review we thoroughly categorize P–N, P–S, and P–C natural organophosphorus compounds. Information on biological source, biological activity, and biosynthesis is included, if known. This review also summarizes the role of phosphorylation on unusual amino acids in proteins (N- and S-phosphorylation) and reviews the natural phosphorothioate (P–S) and phosphoramidate (P–N) modifications of DNA and nucleotides with an emphasis on their role in the metabolism of the cell. We challenge the commonly held notion that nonphosphate organophosphorus functional groups are an oddity of biochemistry, with no central role in the metabolism of the cell. We postulate that the extent of utilization of some phosphorus groups by life, especially those containing P–N bonds, is likely severely underestimated and has been largely overlooked, mainly due to the technological limitations in their detection and analysis.

show abstract

Characterization and cAMP inhibition of a lysyl-(N-ϵ-5′-phospho) adenosyl phosphoamidase in Dictyostelium discoideum

Cited by 13 publications

References 11 publications

Plant nucleoside 5'-phosphoramidate hydrolase; simple purification from yellow lupin (Lupinus luteus) seeds and properties of homogeneous enzyme.

Plant nucleoside 5'-phosphoramidate hydrolase; simple purification from yellow lupin (Lupinus luteus) seeds and properties of homogeneous enzyme.

Adenosine Monophosphoramidase Activity of Hint and Hnt1 Supports Function of Kin28, Ccl1, and Tfb3

Natural Products Containing ‘Rare’ Organophosphorus Functional Groups

Contact Info

Product

Resources

About