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fredag 30 november 2018

Glysiinin biosynteesi 1969 (Harper)

Glysiinin muodostuminen on mielestäni ollut niin monimutkainen  ( verrattuna muihin aminohappoihin) , että olen pitänyt sitä epätodennäköisenä ainoana tapana glysiinin muodostukseen, koska kuitenkin glysiinin tarve on niin suuri. Olisiko se osittain essentielli??

Harper esitti Glysiinin biosynteesin  vuonna 1968 ja  asetan sen vanhan  tekstin tähän. Sivulta
GLYCINE (p.331) Siteeraan englantilasita tekstiä.


This aminoacid is nutritionally nonessential since it can be synthesized by many animals (but not the chick).

A. Deamination and transamination of glycine: CH2(NH2)COOH


Liver and kidney possess a sepcial flavoprotein, glycine oxidase, which oxidatively deaminates glycine to glyoxylic acid  CH(O) COOH

CH2(NH2)COOH  +  2H -> CH(NH)COOH     Iminoacid (Intermediate)
 (Oxidative  reaction)
CH(NH)COOH  + H2O  -> CH(O)COOH + NH3 Glyoxylate
(Deamination reaction)
The deamination is readily reversible.

Transamination of  glyoxylate  using either  glutamine(q), glutamate(e), asparagine(n), aspartate(d) or ornithine(ORN)  as an aminodonor, produces GLYCINE (g) .

GLYCINE is glykogenic and antiketogenic. When glycine labeled with isotopic carbon* in the carbonyl group is fed, the carbon is found in the liver glycogen*,  The amino nitrogen of glycine is exchanged readily with other amino acids.  It has therefore been added in considerable quantities of mixtures of the essentil aminoacids for use in parenteral nutrition.

B. Special functions of GLYCINE


1. Synthesis of heme. 

By the use of labeled glycine* it has been found that the alpha-carbon and nitrogen atoms are used in the synthesis of the porfyrin moiety of hemoglobin. The nitrogen in each pyrrole ring is derived from alfa-nitrogen and an adjoining carbon from the alpha-carbon* of glycine. the alpha-carbon* is also the source of the methylene bridges alpha*, beta*, gamma*, delta*) of the porphyrin structure. For every four glycine nitrogen atoms utilized, eight alpha-carbon* atoms enter the porphyrin molecule.

 2. Formation of Glycine from Choline by way of Betaine 


The oxidation of Choline forms Betaine.
(CH3)3N-CH2-CH2OH, Choline
(CH3)3N-CH2-CHO , Betaine aldehyde
(CH3)3N-CH2-COOH, Betaine 
Betaine is demethylated to Glycine by the loss of  the first methylgroup  in transmethylation, and the last two , by oxidfation to formate.
(CH3)2N-CH2-COOH , Dimethylglycine  and CH3 (transmethylated)
CH3N-CH2-COOH, Sarcosine  and  CH2O  Formaldehyde
NH2-CH2-COOH , Glycine   and CH2O Formaldehyde

3. Conversion of Glycine to Serine


Glycine is readily converted to Serine. Tracer studies  suggest the following mechanism.  (Glycine  gets   "activate Formate-carbon"   and become Serine)
CH2(NH2)COOH  +activated Formate  C*H2O  >  Serine , HOC*H2-CH(NH2)-COOH
The source of the Formate carbon which is added to Glycin to form beta-carbon of Serine has been studied  1956. Mackenzie  has shown that  a 50% yield of Serine  occurs  in the metabolism of Sarcosine by liver mitochondria. Tracer studies  reveal that the beta-carbon of the Serine is derived  from methyl carbon of Sarcosine.  It is suggested , that oxidation of the methyl group produces  "an active one -carbon moiety"  ( "active  Formaldehyde") , which either condenses with Glycine  to form Serine or accumulates as formaldehyde. Labeled formaldehyde   does not participate  in the synthesis of  of Serine in this mitochondrial system
The beta-carbon of Serine serves then as a source of methyl groups for Choline or Thymine synthesis. From the consideration of the above reaction it is apparent that this represents transfer of methyl groups by way of one-carbon(formate) moiety utilizing Glycine and Serine as carrier molecules.  This tyoe oif methyl carbon transfer bas been called "transformalation" by mackenzie.

Huom.  Kirjan kuva esittää Glysiinin konversion Seriiniksi ja myös Seriinistä tiet takasin kohti Glysiiniä, sivu 336:
the conversion of Serine to Glycine involves the loss of the beta-carbon, giving an active one-carbon moiety for methylation or for purine synthesis. If Serine is decarboxylated, ethanolamine is formed.

Sivu 332 Serine decarboxylated   (Missä organismissa tämä 1968  kaava on  pätevä?)
HOCH2-CH2NH2  , ethanolamine

Figure: ethanolamine > Glycolaldehyde and NH3
HOCH2-CHO  ,Glycolaldehyde

Oxidation ov  Glycolaldehyde +   B2 vitamin> Glycolate

HO-CH2-COOH Glycolate oksidation to Glyoxylate HC(O)COOH

Glyoxylate  transamination to Glycine. 

(Siis tässä tulisi Seriinistä Glysiiniä)

4. Synthesis of purines 

The entire Glycine-molecule is utilized to form positions 4, 5 and7 of the purine skeleton.

5. Synthesis of creatine.

The sarcosine (N-methyl-Glycine) component of creatine is derived from Glycine.

 6. A constituent  of Glutathione

The tripeptide glutathione is compound of glutamic acid, cysteine and glycine. The nitrogen in Glutathione is not available for transamination. Note that the peptide bond is with the gamma-COOH of glutamate.

7. Conjugation with Glycine

Glycine conjugates with Cholic acid, forming the bile acid Glycocholic Acid.
With Benzoic acid it forms  hippuric acid ( excreted in the urine).  This reaction is used  as a test of liver function.

8. Oxidation of Glycine

Two pathways for Glycine breakdown have shown,.
One involves  conversion to Serine, the other  formation of Glyoxylate  after deamination by Glycone oxidase.  The further breakdown of Glyoxylate has been studied in rat liver and kidney ( Nakada 1953), where formate ( muurahaishappo, myrsyra)  results from oxidative  decarboxylation  of glyoxylate.
CH(O)COOH +  ½O2 -> HCOOH,  Formate
Formate is oxidiczed  readily by many tissues  to Oxalate
HCOOH + ½O2 -> (COOH)2 , Oxalate ( oksaalihappo, oxalsyra)

Primary hyperoxaluria is a metabolic disease characterized biochemically by continous high urinary  excretion of oxalate which is unrelated to the dietary intake of oxalate (1958).
The history of the disease is that  of progressive bilateral calcium oxalate urolithiasis, nephrocalcinosis, and recurrent infection of urinary tract. (-> renal failure,  hypertension).
The excess ocalate is apparently  of endogenous  origin, possibly from glycine, which, as shown above, after deamination forms glyoxylate, itself a direct source of oxalate.  The metabolic defect in this disease is considered to be a disorder of glyoxylate metabolism associated with failure to convert glyoxylate to formate etc, or to convert it back to glycine by transamination, as shown  above and below. (Fig. above: Gly;  Glyoxylate  ;   CO2  and  Formate
Formate ; Formyltetrahydrofolic acid and  CO2.
Fig. below:  Specific  aminotransaminases)
As a result the excess glyoxylate is oxidized to oxalate.
Two specific aminotransaminases are known to catalyze  conversion of glyoxylate to glycine:
a) Glutamic-glyoxylate-aminotransaminase    ( Glycine  and alfa-ketoglutarate)
b) alanine-glyoxylate aminotransaminase (Glycine and pyruvate) 
In human liver, activity of alanine-glyoxylate aminotransaminase is about 20 times higher than that of the glutamic-glyoxylate aminotransamianse.
  A deficiency of the alanine transaminase together with some impairment of oxidation of glyoxylate to formate may be the biochemical explanation for the inherited metabolic dicease, primary hyperoxaluria.
As might be expected  , vitamin B6-deficient animals excrete markedly increased quantities of oxalate because the glutamic or alanine transaminase reactions are  vitamin B6- dependent.  The excretion of oxalate in B6-deficient rats can be anhanced by feeding glycine or by feeding vitamin B6 antagonists. (1959). However, administration of vitamin B6 has been  benefits in clinical cases of endogenous hyperoxaluria.

(Välillä kommentti: Nykyaikainen  kahvin liika käyttö aiheuttaa vajetta B6 vitamiinissa. Sillä voi olla välillistä vaikutusta  kudosten glysiiniaineenvaihduntaan).

9. The succinate-glycin cycle .

The important role of Glycine in porfyrin and purine synthesis, suggests  a relation of the metabolism of Glycine to Citric acid cycle (1955).These pathways for  the metabolism of  Glycine have summarized in "succinate-glycin cycle".  Succinate ( meripihkahappo), as "active succinate" ( Succinyl-CoA) condenses on  the alpha-carbon*atom of glycine  to form  alfa-amino-beta-ketoadipic acid. It is at this point that the metabolism of Glycine  is  (one way-)linked to the Citric Acid Cycle, which provides  succinyl-CoA. 

(Tässä siis Glysiini  inkorporoituu, osat hyödynnetään    eikä ole.  Glysiinimuoto ole  palautettavissa  vaan osaset joutuvat  lopulta eritystiehen  a) hemoglobiinin  porfyriinirenkaan hajotessa, b) puriinirenkaan hajotessa, c)  ureasykliin karbamyyliryhmän myötä . Sitruunahapposykli ei menetä näitä työkalujäseniään  meripihkahapoa tai   alfaketoglutraattia tässä Glysiinin  työstämisessä). 

 Alpha-amino-betaketoadipic acid  gives away  CO2  (via Biotin, ATP, MG++, glutamine  to  Carbamyl   and to ureacycle).   and  is converted to delta-aminolevulinic acid. This compound serves as a common precursor for porphyrin synthesis aswell as,  after deamination, a carrier molecule ( alfa-ketooglutaraldehyde)  for the introduction of the ureido carbons (2) and (8) into the purine ring.
Succinate and  ketoglutarate, which may return to the citric acid cycle, are also formed. 





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