Methionine methylation reactions

It is important in the body as, except for methionine, it is the only substance known to take part in methylating reactions. Sometimes regarded as a member of the vitamin B group. 

The amino acid methionine is formed by a melhylation reaction of homo cysteine with iV-methyltetrahydrofolate. The stereochemistry of the reactior has been probed by carrying out the transformation using a donor with a "chiral methyl group" that contains protium (H), deuterium (D), and tritium (T isotopes of hydrogen. Does the methylation reaction occur with inversion oi retention of configuration ... 

In this reaction sequence acetic acid synthesis requires methyl transfer as CH3 to a Co(I)-corrin by N2 5-methyltetrahydrofolate monoglutamate to give a methylcorrinoid intermediate which is carboxylated to give a carboxymethylcorrinoid. This carboxymethylcorrinoid would then be reductively removed by NADPH to give acetic acid and regenerate the Co(I)-corrin. In contrast to the methyl-transfer proposed for the methionine synthetase reaction, this mechanism suggests that CH3-stabilized by Co attacks CO2 to give a carboxymethylcorrinoid intermediate. 

A high intracerebral level of S-adenosylhomocysteine may inhibit methylation reactions involving S-adenosyl-methionine. The metabolic repercussions would be extensive, including deficient methylation of proteins and of phos-phatidylethanolamine as well as an inhibition of catechol-O-methyltransferase and histamine-N-methyltransferase. 

SMM synthesis is mediated by the enzyme methionine S-methyltransferase (MMT) through the essentially irreversible, AdoMet-mediated methylation of methionine.48"5 Both MMT and SMM are unique to plants 48,50 The opposite reaction, in which SMM is used to methylate homocysteine to yield two molecules of methionine, is catalyzed by the enzyme homocysteine S-methyltransferase (HMT).48 Unlike MMT, HMTs also occur in bacteria, yeast, and mammals, enabling them to catabolize SMM of plant origin, and providing an alternative to the methionine synthase reaction as a means to methylate homocysteine. Plant MMT and HMT reactions, together with those catalyzed by AdoMet synthetase and AdoHcy hydrolase, constitute the SMM cycle (Fig. 2.4).4... 

It is the role of jV5-methyl THF which is key to understanding the involvement of cobalamin in megaloblastic anaemia. The metabolic requirement for N-methyl THF is to maintain a supply of the amino acid methionine, the precursor of S-adenosyl methionine (SAM), which is required for a number of methylation reactions. The transfer of the methyl group from jV5-methyl THF to homocysteine is cobalamin-dependent, so in B12 deficiency states, the production of SAM is reduced. Furthermore, the reaction which brings about the formation of Ns-methyl THF from N5,N10-methylene THF is irreversible and controlled by feedback inhibition by SAM. Thus, if B12 is unavailable, SAM concentration falls and Ah -methyl THF accumulates and THF cannot be re-formed. The accumulation of AT-methyl THF is sometimes referred to as the methyl trap because a functional deficiency of folate is created. 

In biological methylation, the 5-methyl group of the amino acid L-methionine is used to methylate suitable O, N, S, and C nucleophiles. First, methionine is converted into the methylating agent S-adenosylmethionine (SAM). SAM is nucleoside derivative (see Section 14.3). Both the formation of SAM and the subsequent methylation reactions are nice examples of biological Sn2 reactions. 

The coenzyme tetrahydrofolate (THF) is the main agent by which Ci fragments are transferred in the metabolism. THF can bind this type of group in various oxidation states and pass it on (see p. 108). In addition, there is activated methyl, in the form of S-adenosyl methionine (SAM). SAM is involved in many methylation reactions—e. g., in creatine synthesis (see p. 336), the conversion of norepinephrine into epinephrine (see p. 352), the inactivation of norepinephrine by methylation of a phenolic OH group (see p. 316), and in the formation of the active form of the cytostatic drug 6-mercaptopurine (see p. 402). 

Figure 7.64 The role of methionine in methylation reactions and the mechanisms underlying ethionine hepatotoxic-r. ity. After the substrate is methylated, the S-adenosyl homocysteine remaining is broken down into homocysteine and adenine, both of which are reused. When S-adenosyl ethionine is formed, however, this recycling is reduced (=), and a shortage of adenine and hence ATP develops.

Another example of leaving group activation is the utilization of S-adenosyl-methionine rather than methionine in methylation reactions. A relatively basic thiolate anion has to be expelled from methionine, while the nonbasic neutral sulfur is displaced from the activated derivative (equation 2.68) ... 

With methyl iodide as the alkylating reagent, Link and Stark (133) prepared a monosubstituted sulfonium salt. Methionine 29 was strongly indicated as the principal site of modification. The activity of the derivative toward C>p was the same as RNase-A, and the methylation reaction was not affected by competitive inhibitors such as 2 (3 )-UMP. 

A large number of both endogenous and exogenous compounds can be methylated by several N-, 0-, and S-methyl transferases. The most common methyl donor is S-adenosyl methionine (SAM), which is formed from methionine and ATP. Even though these reactions may involve a decrease in water solubility, they are generally detoxication reactions. Examples of biologic methylation reactions are seen in Figure 7.18. 

Homocysteine is formed as an intermediary amino acid in the methionine cycle (Fig. I). Methionine is metabolized to s-adenosylmethionine (SAM), the methyl donor in most methylation reactions and essential for the synthesis of creatinine, DNA, RNA, proteins, and phospholipids. SAM is converted by methyl donation to s-adenosylhomocysteine (SAH), which is then hydrolyzed to homocysteine. SAH is an inhibitor of methyl group donation from SAM. 

Indolmycin.—It has been shown that the C-methyl group in indolmycin (82) originates from the methyl group of methionine with inversion of configuration (cf. Vol. 8, p. 23). Previously published in preliminary form,71 the results are now available in a full paper.72 In addition, it has been shown that the A-methylation reaction which occurs in the course of the biosynthesis of indolmycin (82) also proceeds with inversion of configuration of the methyl group of methionine. Similar methyl-transfer with inversion has been recorded in the catechol-O-methyl-transferase reaction,73 and in this case it has been concluded that there is a tight SN2 transition state for the methyl-transfer.74... 

Figure 21-3. The methionine synthase reaction. Methionine synthase catalyzes the remethylation of homocysteine to methionine. In the first half reaction (1), a methyl group is transferred from 5-methyl tetrahydrofolate (5-MTHF) to the reduced form of cobalamin [Cob(I)], generating methyl-cobalamin [Methyl-Cob(III)] and tetrahydrofolate (THF). During the second half reaction (2), the methyl group is transferred from methylcobalamin to homocysteine, generating methionine. During the catalytic reaction, Cob(I) occasionally becomes oxidized, producing an inactive form of cobalamin, cob(II)alamin [Cob(II)]. The enzyme methionine synthase reductase (MTRR) then reactivates Cob(II) through reductive methylation, producing methyl-Cob(III). SAM, 5-adenosylmethionine SAH, 5-adeno-sylhomocysteine.

The most widely used methyl donor for enzymatic methyl transfer is the cofactor S-adenosyl-L-methionine (SAM). The methyl moiety on the L-methionine is supplied by another known methyl donor, N5-methyl tetrahydrofolate.30 To date, numerous enzymes that perform SAM- dependent methylation reactions have been described in plants, and several reports attempting to sort out their evolutionary relationships have been published.31- 3... 

Phosphatidylserine arises by an exchange of the ethanolamine residue of phosphatidylethanolamine for a seryl group. Decarboxylation of the serine of phosphatidylserine reforms phosphatidylethanolamine. Three successive methylation reactions convert phosphatidylethanolamine to phosphatidylcholine. 5-Adenosyl-methionine is the methyl-group donor (Chap. 15) (see Fig. 13-14). 

The mammalian synthesis of methionine is more complex and requires cobalamin, a coenzyme form of vitamin B12. Note that because methionine is an essential amino acid, it must be supplied in the diet methionine that is used for methylation (Fig. 15-20) is degraded to homocysteine, and this is remethylated to give methionine. These reactions merely recycle methionine and do not constitute a means of net synthesis. 

With the assumption that reticulines are also precursors in mammalian synthesis of morphine, it was challenging to investigate whether they could be produced by enzymatic reactions similar to those utilized in benzylisoquinoline-producing plants (274). This plan focused attention on reactions controlled by the enzyme catechol 0-methyltransferase (COMT), using 5-adenosyl-L-methionine (SAM) for the methylation reaction. Mammalian COMT is present in mammalian tissues, particularly the liver, and an enzyme preparation from rat liver was used for the experiments. It was found that (S)-norcoclaurine, which is the first isoquinoline produced in benzylisoquinoline-producing plants, was similarly O-methylated in vitro by SAM in the presence of COMT, and a reverse proportion of methylated products was obtained with the (/ )-enantiomer (277). Similar 0-methylation of (5)-4 -demethylreticuline (3 -hydroxy-N-methylcoclaurine), prepared by total synthesis (162), however, afforded almost exclusively (5)-orientaline, with a methoxy group at C-3 and not at C-4 as in (5)-reticuline (Fig. 37) (762). 

Methylcobalamin is completely different from adenosylcobalamin because it is essentially a conduit for synthetic reactions catalyzed by methyltransferases, illustrated in Scheme 2 for the case of methionine. These reactions depend on the supemucle-ophilicity of cob(I)alamin. In one case, this species removes a methyl group from A -methyltetrahydrofolate with the formation of methylcobalamin, and then transfers this group to the acceptor homocysteine, which results in the synthesis of methionine (see Scheme 2). 

Although methylation reactions are mainly concerned with endogenous substrates, a number of drugs are also methylated [Eq. (33)] by non-specific methyltrans-ferases that are present in the liver, lung, and other tissues. The enzymes are located mostly in the cytosol, but they may also be membrane-bound. The principal methyl donor is S-adenosylmethionine (SAM) which is formed from ATP and L-methionine. 

S-adenosyl-L-methionine (SAM)-dependent methyl-ation was briefly discussed under Thiomethylation (see Figure 14). Other functional groups that are methylated by this mechanism include aliphatic and aromatic amines, N-heterocyclics, monophenols, and polyphenols. The most important enzymes involved in these methylation reactions with xenobiotics are catechol O-methyltransferase, histamine N-methylt-ransferase, and indolethylamine N-methyltransferase - each catalyzes the transfer of a methyl group from SAM to phenolic or amine substrates (O- and N-methyltransferases, respectively). Methylation is not a quantitatively important metabolic pathway for xenobiotics, but it is an important pathway in the intermediary metabolism of both N- and O-contain-ing catechol and amine endobiotics. 

Once inside the cell, folates participate in a number of interconnected metabolic pathways involving (1) thymidine and purine biosynthesis necessary for DNA synthesis, (2) methionine synthesis via homocysteine remethylation, (3) methylation reactions involving S-adenosylmethionine (AdoMet), (4) serine and glycine interconversion, and (5) metabolism of histidine and formate (see Figure 8). Via these pathways. 

The reaction was applied to model dipeptides exemplified by the y-phenyl-hydrazide of N-carbobenzoxy-a-L-glutamyl-L-methionine methyl ester (4) and found to afford the carboxylic acid (S) in good yield without disturbance of the carbo-benzoxy and ester protective groups. The results suggest use of the phenylhydrazide group for protection of carboxyl groups in peptide chemistry. 

---