Monday, April 9, 2012

Glutamate




General features
 R = -(CH2)2COO- at cytoplasmic pH (pH~7)
 pKa (terminal carboxyl group)= 4.25

Physiological roles
            In prokaryotes, an important function is to donate its alpha-amino group to other amino acids that are being synthesized:

Each transaminase enzyme is specific for the amino acid that will be aminated (or deaminated).  Details of the mechanism can be found here:  http://www.bmb.leeds.ac.uk/illingworth/bioc1010/index.htm

            Another function of L-glutamate is to maintain regular L-glutamine levels while assimilating nitrogen in one of two pathways (to be discussed below and in the next post). Also, it can be fermented in various pathways under anaerobic conditions (X).
            A structural function is that its enantiomer, D-glutamate is typically a member of the tetrapeptide bridge in peptidoglycan linkages:
           
L-glutamate is first added to L-alanine, then glutamate racemase (murI) will convert it to the D enantiomer.
            Another notable structural function of D-glutamate is in the poly-D-glutamyl capsule of Bacillus anthracis, the capsule being one of its main virulence factors (avoidance of phagocytosis). More on this requirement for B. anthracis pathogenicity can be found: http://www.nature.com/emboj/journal/v24/n1/full/7600495a.html#B25

Synthesis
            Like many of the amino acids, L-glutamate corresponds to a ‘parent’ metabolite of the TCA cycle, α-ketoglutarate, whose supply can be increased or decreased depending on the relative need for glutamate and its downstream products. When α-ketoglutarate derivatives are highly demanded, they will deplete its supply and are thus cataplerotic, taking TCA carbons from the cycle. When α-ketoglutarate derivatives are used to increase its concentration because some other portion of the TCA cycle requires concentrations of its intermediates (say, an increase demand for fumarate for tyrosine synthesis), the conversion of glutamate to α-ketoglutarate would be anaplerotic, because it would be adding carbons to the TCA cycle.


           
L-glutamate can be regenerated by two significant enzmes, either from existing pools of glutamine + α-ketoglutarate (glutamate synthase, GS) or via NH4+ + α-ketoglutarate (glutamate dehydrogenase, GOGAT. In the synthesis of glutamate, this enzyme seems to be named backwards).


       

The pathway that uses glutamate synthase (GOGAT) is often coupled with glutamine synthase (GS) for the constant recycling of glutamate/glutamine. The second pathway that uses glutamate dehydrogenase (GDH) is not used as often because the the enzyme itself favors an equilibrium more toward the ammonium, α-ketoglutarate products due to its high Km for NH3 (meaning it needs a higher [NH3] to bind this substrate). Use of this enzyme under normal [NH3] would then lead to an excess of un-assimilated/free ammonia within the cell which is toxic. Also of interest is the fact that GDH does not use an ATP for the catalysis of its reaction. So generally speaking, under high [NH3] and low energy levels the bacterial cell will use glutamate dehydrogenase. The glutamate/glutamine pair, which differ by an amino group, is used for nitrogen assimilation (as will be described in the next post). Given this difference, it is convenient for the cell to continuously cycle them to maintain a constant level of intracellular ammonia.


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