Purification and Properties of Threonine Synthetase of Neurospora

Flavin, Martin; Slaughter, Clarence

doi:10.1016/s0021-9258(18)69487-6

Cited by 70 publications

(15 citation statements)

References 21 publications

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“…C7H16N05P*0.4 H 2 0 requires C, 36.2; H, 7.3; N, 6.0%); 6H(300 MHz; D20) 1.32, 1.33 (2 x t, 3 H, .86 mmol) in dry dichloromethane (20 ml) was treated with trifluoroacetic acid (4 ml) and stirred at room temperature overnight. The mixture was concentrated, azeotroped with toluene, and chromatographed on a 4 mm Chromatotron plate (20% Pr'OH-EtOAc) to afford 0.91 g (71%) of the lactone(19) as a colourless oil (Found: C, 52.0; H, 6.0; N, 3.9. MHz; CDC1,)1.32 (t, 3 H, J 7.0 Hz), 1.56, 1.58, 1.61 (3 x d, 3 H, J 14.0 Hz), 2.20-3.00 (m, 2 H), 4.02-4.22 (m, 2 H), 4.384.72 (m, 2 H), 5.11 (m, 2 H), 5.91, 5.99, 6.05, 6.30 (4 x br d, 1 H, J 7.2 Hz), and 7.34 (m, 5 H); 6,(40.3 MHz; CDCl,) 45.5 (0.33P), 47.6 (0.33P), and 48.0 (0.34P); m/z (FAB MS)…”

mentioning

confidence: 99%

Design and synthesis of γ-oxygenated phosphinothricins as inhibitors of glutamine synthetase

Walker¹,

McDonald²,

Franz³

et al. 1990

J. Chem. Soc., Perkin Trans. 1

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The ability of L-y-hydroxyglutamic acids to act as substrates of the enzyme glutamine synthetase (GS) was exploited as a rationale for the synthesis of y-oxygenated analogues of the naturally occurring GS inhibitor phosphinothricin (PPT), The potent new inhibitor DL-Y-hydroxyphosphinothricin (GHPPT) was prepared via a key reaction involving the silicon-mediated addition of ethyl methylphosphinate to benzyl 2-benzyloxycarbonylamino-4-oxobutyrate. The resulting intermediate was also converted to various derivatives useful in probing structure-activity relationships in the GH PPT series. The y-oxygenated phosphinothricins display enzyme inhibitory activity as well as in vivo phytotoxicity.

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mentioning

confidence: 99%

Design and synthesis of γ-oxygenated phosphinothricins as inhibitors of glutamine synthetase

Walker¹,

McDonald²,

Franz³

et al. 1990

J. Chem. Soc., Perkin Trans. 1

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“…Significantly, throughout the purification process, His 6 -RhiB exhibited a yellow color characteristic of either a bound flavin or PLP cofactor. Threonine synthases characterized to date employ the use of PLP as a cofactor to convert O -phospho- l -homoserine into threonine. ,, UV–vis spectroscopy of purified His 6 -RhiB revealed the presence of an absorption maximum at 420 nm characteristic of PLP-containing enzymes similar to other characterized threonine synthases (Supplementary Figure S1). We next sought to determine the ability of this enzyme to convert PHSer into threonine.…”

Section: Resultsmentioning

confidence: 90%

Molecular Basis of Bacillus subtilis ATCC 6633 Self-Resistance to the Phosphono-oligopeptide Antibiotic Rhizocticin

et al. 2019

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Rhizocticins are phosphono-oligopeptide antibiotics that contain a toxic C-terminal (Z)-L-2amino-5-phosphono-3-pentenoic acid (APPA) moiety. APPA is an irreversible inhibitor of threonine synthase (ThrC), a pyridoxal 5'-phosphate (PLP)-dependent enzyme that catalyzes the conversion of O-phospho-L-homoserine to L-threonine. ThrCs are essential for the viability of bacteria, plants, and fungi and are a target for antibiotic development, as de novo threonine biosynthetic pathway is not found in humans. Given the ability of APPA to interfere in threonine metabolism, it is unclear how the producing strain B. subtilis ATCC6633 circumvents APPA toxicity. Notably, in addition to the housekeeping APPA-sensitive ThrC (BsThrC), B. subtilis encodes a second threonine synthase (RhiB) encoded within the rhizocticin biosynthetic gene cluster. Kinetic and spectroscopic analyses show that PLP-dependent RhiB is an authentic threonine synthase, converting O-phospho-L-homoserine to threonine with a catalytic efficiency comparable to BsThrC. To understand the structural basis of inhibition, we determined the crystal structure of APPA bound to the housekeeping BsThrC, revealing a covalent complex between the inhibitor and PLP. Structure-based sequence analyses reveal the molecular determinants within the RhiB active site responsible for rendering this ThrC homolog resistant to APPA. Together, this work establishes the self-resistance mechanism utilized by B. subtilis ATCC6633 against APPA exemplifying one of many ways by which bacteria can overcome phosphonate toxicity.

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“…Polarity with respect to flanking markers is not reversed when the met-6 region is inverted relative to the centromere (722). (124,208,209,351,352,518, 547, %5). For conversion of threonine to isoleucine, see Fig.…”

Section: In(il -) Ir)h4250mentioning

confidence: 99%

“…17. Biosynthetic pathways of homoserine, threonine, and methionine, showing sites of gene action(124,208,209,351,352,518, 547, %5). For conversion of threonine to isoleucine, see Fig 15…”

mentioning

confidence: 99%

Chromosomal loci of Neurospora crassa

Perkins¹,

Radford²,

Newmeyer³

et al. 1982

Microbiol Rev

259

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I I ' ~~~~T h Ñ CiOr(r r N z ADDITIONAL LOCI IN LINKAGE GROUP ILinked to mt (8 to 12%) Linked to mt, acr-3 (5%) Right of arg-i (5/29) Linked to so and aro-8 Right of un-18 Between ad-5 and the centromere Between arg-i (4%) and his-3 (I to 2%) Between In(H4250) and T(39311) Left of mt (23%) Linked to mt (2%) and aza-i (39%o) Left of nit-2 (30%) Left of mt (14%) Linked to un-i (9%), probably to the right Linked to hom (1%), probably to the right Linked to mt (17 to 22%) Between T(NM103) and al-2 (18%) Between T(4540) and cr-2Between thi-i (12 to 20%/6) and ad-9 (5%) Linked to ad-5 (1/54), probably to the left Linked to mt (6%) Linked to cys-5 (<1%), probably to the right Right of ad-9 (12%), linked to al (0/76) Right of his-3 (2%) Between the T(AR173) breakpoints Linked to al-2 (10%) and nic-i (1 to 5%) Linked to ad-9 (0/44); right of thi-i (2%) Linked to mt (5%), probably to the left Linked to al-I (10%) Right of os-i (3%) Between nit,i (5 to 19%) and al-I (6 to 19%io) Linked near arg-i Left of al-2 (25%o) Left of mt (16%) Right of nic-2 (6%) Linked to ad-9 (2%), probably to the right Left of cr-i (15%) Left of mt (10%) Probably between his-2 (3%) and cr-i (3%) Between T(NM103) and the right tip Between mt (20%) and arg-i (1%) Between ad-3 (6%) and al-i (16%) Between his-3 and nic-2 Between cyh-i (5%) and al-I (7%) Linked to cyh-i (18%), al-i (2%), and mb-2 (6%) Between arg-i (3%) and T(39311) Between tre (41%) and ad-9 Linked to mt (9%) Linked to mt (20%) Complex phenotype. Alternative requirements: 2-acetylaminofluorine, certain azo dyes, or certain single amino acids. Cold sensitive. Originated among progeny of a rib-i strain that had become tolerant to 2-acetylaminofluorine.

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Purification and Properties of Threonine Synthetase of Neurospora

Cited by 70 publications

References 21 publications

Design and synthesis of γ-oxygenated phosphinothricins as inhibitors of glutamine synthetase

Design and synthesis of γ-oxygenated phosphinothricins as inhibitors of glutamine synthetase

Molecular Basis of Bacillus subtilis ATCC 6633 Self-Resistance to the Phosphono-oligopeptide Antibiotic Rhizocticin

Chromosomal loci of Neurospora crassa

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