Phosphoribosyl AMP

This enzyme [EC 1.1.1.23] catalyzes the reaction of l-histidinol with two NAD+ to produce L-histidine and two NADH. L-Histidinal will also serve as a substrate for this protein. The Neurospora crassa enzyme will also catalyze the reactions of phosphoribosyl-AMP cyclohydrolase [EC 3.5.4.19] and phosphoribosyl-ATP pyrophosphatase [EC 3.6.1.31]. 

This enzyme [EC 3.5.4.19] catalyzes the hydrolysis of 5-phosphoribosyl-AMP to produce 5-(5-phospho-D-ribo-sylaminoformimino)-l-(5-phosphoribosyl)tmidazole-4-carboxamide. The enzyme isolated from Neurospora crassa can also catalyze the reactions of histidinol dehydrogenase and phosphoribosyl-ATP pyrophosphatase. 

Fig. 2. The pathway of histidine biosynthesis. Enzymes a, ribosephosphate pyrophos-phokinase E.C. 2.7.6.1 b, ATP-phosphoribosyltransferase, E.C. 2.4.2.17 c, phosphoribosyl-AMP cyclohydrolase, E.C. 3.5.4.19 d, N-(5 -phospho-D-ribosylforminino)5-amino-l-(5"-phos-phoribo yl)-4-imidazole carboxamide isomerase, E.C. 5.3.1.16 e, glutamine amidotransferase f, imidazolglycerolphosphate dehydratase E.C. 4.2.1.19 g, histidinol-phosphate aminotransferase E.C. 2.6.1.9 h, histidinol phosphatase, E.C. 3.1.3.15 i, histidinol dehydrogenase, E.C. 1.1.1.23.

Figure 5.82 Biosynthesis of histidine. PRPP, 5-phosphoribosyl-a-l-pyrophosphate PRATP, AT-5 -phosphoribosyl-ATP PRAMP, M-S -phosphoribosyl-AMP 5 -ProFAR, yV -[(5. phosphoribosyl)-formimino]-5-aminoiinidazole-4-carboxamide-ribonucleotide 5 -PRFAR, A/ -[(5 -phosphoribulosyl)-formimino]-5-aminoimidazole-4-carboxainide-ribonuc leotide IMGP, imidazole glycerol-phosphate AICAR, 5 -phosphoribosyl-4-carboxamide-5-aminoimidazole lAP, imidazoleacetol-phosphate HOL-P, L-histidinol-phosphate HOL, L-histidinol HAL, L-histidinal.

We have attempted to dissect the incorporation of NAD into these two reactions by snake venom phosphodiesterase (SVPDE) digestion. The rate of incorporation of label into 5 -AMP gives an estimate of the rate of initiation and the rate of incorporation into phosphoribosyl-AMP(PR-AMP) estimates the rate of elongation. AE-cellulose chromatography can be used to monitor the proportion of 5 -AMP deriving from mono- and poly(ADP-ribose) residues respectively. This provides an important check since, in permeabilised cells, mono(ADP-ribose) residues can be produced other than in the nucleus and also by non-enzymic reactions [9],... 

Properties of p21.bot. We have used pure C3 to further characterize the p21.bot ADP-ribosylation free of interference from actin labeling. Upon complete digestion with snake venom phosphodiesterase, p P]ADP-ribosylated p21.bot releases [ P]AMP but not [ P]phosphoribosyl AMP, indicating that C3 catalyzes a mono-ADP-ribosylation. 

Abbreviations used PRPP, phosphoribosylpyiophosphate PR-ATP, phosphoribosyl-ATP PR-AMP, phosphoribosyl-AMP PR-AIC, 4-amino-S-iinidizolecarboxainide ribonucleotide ribosyl-AIC, 4-amino-S-imidazoIecarboxamide ribonucleoside TRA, 1,2,4-triazole-3-alanine. 

ATP phosphoribosyltransferase 2 phosphoribosyl-ATP pyrophosphatase 3 phosphoribosyl-AMP cyclohydrolase 4 isomerase 5 formyltransferase, inosine monophosphate cyclohydrolase 6 adenylosuccinate synthetase 7 adenylosuccinate lyase 8 adenylate kinase 9 imidazole glycerol phosphate dehydratase 10 histidinol phosphate aminotransferase 11 histidinol phosphatase 12 histidinol dehydrogenase... 

Fig. 4. Histidine tnosynthess. Intermediates PRPP, phosphoribosyl pyrophosidiate PR-ATP, JV-phoqjhoribosyl-ATP PR-AMP, A/ -phosphoribosyl-AMP PR-FAIC-RN, phosphoribosyl for-mimino-S-amino imidazole carboxamide ribonucleotide PRu-FAIC-RN, phosphoribulosyl for-mimino-5-aniino imidazole carboxamide ribonucleotide IGP, imidazole glycerol phosphate lAP, imidazole acetol idrosphate HP, histidinol (dioqrhate HOL, histidinol. Enzymes (I) ATP phos-phoribosyltransferase (EC 2.4.2.17) (2) phosphoribosyl-ATP pyrophosphohydrolase (EC 3.6.1.31) (3) idiospharibosyl-AMP cyciohydrolase (EC 3.5. A19) (4) PR-FAIC-RN isomerase (EC S.3.1.16) (S) glutamine amidotransferase (6) cyclase (7) imidazole glycerol phosphate dehydratase (EC 4.2.1.19) (8) histidinol-phosphate aminotransferase (EC 2.6.1.9) (9) histidinol phosphatase (EC 3.1.3.15) (10) histidinol dehydrogenase (EC 1.1.1.23).

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