Aspartate family synthesis

Although L-phenylalanine is a protein amino acid, and is known as a protein acid type of alkaloid precursor, its real role in biosynthesis (providing C and N atoms) only relates to carbon atoms. L-phenylalanine is a part of magic 20 (a term deployed by Crick in his discussion of the genetic code) and just for this reason should also be listed as a protein amino acid type of alkaloid precursor, although its duty in alkaloid synthesis is not the same as other protein amino acids. However, in relation to magic 20 it is necessary to observe that only part of these amino acids are well-known alkaloid precursors. They are formed from only two amino acid families Histidine and Aromatic and the Aspartate family . 

The aspartate family also contains asparagine, lysine, methionine, and threonine. Threonine contributes to the reaction pathway in which isoleucine is synthesized. The synthesis of isoleucine, often considered to be a member of the pyruvate family, is discussed on p. 467. 

The synthesis of the other members of the aspartate family (Figure 14.8) is initiated by aspartate kinase (often referred to as aspartokinase) in an ATP-requiring reaction in which the side chain carboxyl group is phosphorylated. Aspartate... 

Since the substrate specificity of individual aminotransferases may vary widely, it is not known whether the nitrogen atoms of the aspartate family and branched-chain amino acids are derived from a single, or from multiple precursors (Table I). Utilization of a common amino donor in aminotransferase catalyzed reactions would strengthen the biosynthetic relationship among the pathway products, whereas multiple precursors could tend to balance the synthesis of these amino acids with that of other protein precursors in a type of crosspathway or interfamily regulation. 

An alternative approach to estimating the metabolic capabilities of chloroplasts entails measurement of the light-dependent metabolism of radioactive tracers. Using isolated pea chloroplasts. Mills and Wilson (1978a) found that lysine, methionine, threonine, and isoleucine were synthesized from [ C]aspartate. Further evidence that aspartate was being metabolized via the anticipated pathways was provided by the demonstration that the synthesis of homoserine was inhibited by lysine and threonine (Lea et al., 1979). These results, combined with those relating to enzyme localization, lead to the concept that chloroplasts contain a complete functional sequence of enzymes which can facilitate the synthesis of the aspartate family and at least some of the branched-chain amino acids. This is consistent with the importance of chloroplasts in ammonia assimilation (Miflin and Lea, this volume. Chapter 4) and with the evidence that protein can be synthesized from CO2 in isolated plastids (Shepard and Leven, 1972 Huberer al., 1977). The actual fraction of [ ]02 which is utilized for amino acid biosynthesis in isolated plastids is usually quite small. Thus, reactions which normally occur outside of chloroplasts are considered to be of major importance in the synthesis of carbon skeletons such as oxaloacetate or pyruvate (Kirk and Leech, 1972 Leech and Murphy, 1976). 

Fig. 7. Sequential control of the synthesis of the aspartate family of amino adds. Temporal control of the flow of carbon is illustrated by the successive Figs. 1-5. Potential quantitative changes in flow are approximated by the thickness of the solid arrows. As the concentration of an end product is increased (indicated by closed boxes), the pattern of synthesis is altered by utilization of negative (-) or positive (+) regulatory mechanisms as described in the text. The pattern of control which is illustrated assumes that aspartate kinase is sensitive to inhibition only by lysine. Variations of this pattern are discussed in the text.

Regulation of the synthesis of the branched-chain amino acids, like that of the aspartate family, can be viewed in a temporal framework (Fig. 8). However, the nature of the controls associated with the pathway enzymes do not necessarily suggest an obligatory sequence of regulatory interactions. The sequence illustrated in Fig. 8 assumes that each of the end-products would initially be synthesized from its respective precursors. As isoleucine biosynthesis is reduced by inhibition of threonine dehydratase, the competition between pyruvate and 2-oxobutyrate for the active site of acetohydroxyacid synthase would be diminished. This could result in an increased rate of synthesis of leucine and valine (Fig. 8, 2). Leucine would eventually inhibit isopropylmalate synthase and, to a lesser extent, acetohydroxyacid synthase (Fig. 8, 3). The reduced flow of carbon through the pathway would be utilized for the synthesis of valine. As the concentration of valine increased, the activity of acetohydroxyacid synthase would be sharply curtailed due to... 

The sequences of biochemical transformations involved in the synthesis of the aspartate family and branched-chain amino acids in multicellular plants are similar to those that occur in microorganisms. Support for this conclusion has been derived principally from isolation of a number of the requisite enzymes. Information on the kinetic and physical properties of enzymes is best achieved after extensive purification. In contrast, useful predictions of the physiological function of regulatory enzymes depend upon effective enzyme extraction and complete preservation of native properties. Since the latter objective has been emphasized during most investigations of enzymes associated with amino acid biosynthesis in plants, the bulk of our knowledge has been obtained from comparatively crude enzyme preparations. Results of both direct and competitive labeling experiments have added demonstrations of many of the predicted precursor-product relationships and a few metabolic intermediates have been isolated from plants. The nature of a number of intermediate reactions does, however, remain to be clarified notably, the reactions associated with the conversion of dihydropicolinate to lysine and those involved in the synthesis of leucine from 2-oxoisovalerate. 

Synthesis of the Aspartate Family and Branched-Chain Amino Acids J. K. Bryan... 

Bryan, J. K. Synthesis of the aspartate family and branched-chain amino acids. In The Biochemistry of Plants, Vol. 5, Amino Acids and Derivatives (B. J. Miflin, ed.), pp. 403-452. Academic Press, New York 1980... 

Fig. 157. Fine regulation by isoenzymes biosynthesis of the aspartate family of amino acids. ER = endproduct repression, EH = endproduct inhibition. The synthesis of methionine is also subject to-feedback mechanisms, which are, however, now shown here (cf. also Fig. 156).

Enzymes of the pepsin family rarely catalyze the hydrolysis of esters, with the exceptions of, for example, esters of L-/3-penicillactic acid and some sulfinic acid esters. Under suitable conditions, i. e., low pH, high enzyme concentration, and formation of an insoluble peptide, aspartic peptidases are able to catalyze the synthesis of peptides [71] [72],... 

Sulfate Must Be Reduced to Sulfide before Incorporation into Amino Acids The Aspartate and Pyruvate Families Both Make Contributions to the Synthesis of Isoleucine... 

The aspartate (oxaloacetate) family of amino acids includes aspartate, asparagine, methionine, lysine, threonine, and isoleucine (see fig. 21.1). The pyruvate family includes alanine, valine, leucine, and also lysine and isoleucine (see fig. 21.1). Threonine is a precursor of isoleucine. It is converted into isoleucine by a group of enzymes that are also used in the synthesis of valine (fig. 21.10). 

The aspartate and pyruvate families together contain 11 amino acids. Because of the reactions involved in its synthesis, isoleucine is considered a member of both families. Isoleucine and valine use four enzymes in common in their biosynthetic pathways. 

A few steps convert inosinate into either AMP or GMP (Figure 25,9). Adenylate is synthesized from inosinate by the substitution of an amino group for the carbonyl oxygen atom at C-6. Again, the addition of aspartate followed by the elimination of fumarate contributes the amino group. GXP, rather than ATP, is the phosphoryl-group donor in the synthesis of the adenylosuccinate intermediate from inosinate and aspartate. In accord with the use of GTP, the enzyme that promotes this conversion, adenylsuccinate synthase, is structurally related to the G-protein family and does not contain an ATP-grasp domain. The same enzyme catalyzes the removal of fumarate from adenylosuccinate in the synthesis of adenylate and from 5-aminoimidazole-4-jV-succinocarboxamide ribonucleotide in the synthesis of inosinate. 

On the basis of the similarities in their synthetic pathways, the amino acids can be grouped into six families glutamate, serine, aspartate, pyruvate, the aromatics, and histidine. The amino acids in each family are ultimately derived from one precursor molecule. In the discussions of amino acid synthesis that follow, the intimate relationship between amino acid metabolism and several other metabolic pathways is apparent. Amino acid biosynthesis is outlined in Figure 14.4. 

Seven enzyme catalyzed reactions are required for the synthesis of lysine from pyruvate and aspartate semialdehyde as illustrated in Fig. 3. However, enzymes catalyzing only the first and last of these reactions have been isolated from plants. Dihydropicolinate synthase facilitates the condensation of the precursors during a reaction which presumably proceeds in two steps. A double bond between the C-4 of the semialdehyde and the methyl carbon of pyruvate would be formed, with the loss of water, followed by spontaneous ring closure and the loss of a second molecule of water. Catalysis of this reaction in plant extracts was first demonstrated by Cheshire and Miflin (1975) using maize seedlings as the source of the enzyme. Mazelis et al. (1977) detected the enzyme in extracts obtained from six different taxonomic families of plants and partially purified the enzyme from wheat germ. Only the L isomer of aspartate semialdehyde was active as a substrate of this enzyme and strong cooperativity was noted when the concentration of pyruvate was varied. A dihydropicolinate synthase has also been isolated from carrot cells (Matthews and Widholm, 1978). 

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