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Can cells be grown in exogenous supply of Malonyl-CoA and Malate? Need Help
#1
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i am new to this site. so dont know if it is the relevant thread to post my question. Can cells be grown in exogenous supply of Malonyl-CoA and Malate. These are the first product/precursor of the first product of two major pathways and i want to check the effect of inhibition of these two pathways. please someone throw some light here. help would be highly appreciated.
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#2
Dear Billamin,
Welcome to the site ! You can post all your queries related to biotechnology & other biological science in this forum.

As you may be know that Malonyl-CoA is the precursor for the de novo synthesis and elongation of fatty acids chains. Also these are used in formation of flavonoids,stilbenoids, isoflavonoids etc and in many other malonylated compounds, including d-amino acids, malonylated 1-aminocyclopropane-1-carboxylic acid, and flavonoid glycosides.

The answer of your query "Can cells be grown in exogenous supply of Malonyl-CoA and Malate", is No if they are gram negative organisms (cells)or any cells with more lipid content in their membranes.

Since Malate are anions (here consider it in terms of polarity - as we are thinking of exogenous supply-ONLY) and are unable to pass through the lipid bilayer of membranes and hence require carriers (eg 2-oxoglutarate/malate carrier protein)for the same.Therefore if you are thinking of cell with more lipid membrane example (Gram Negative microorganisms ) this may not be possible without carriers and if the only source of growth is this.
Now if you are thinking of exogenous supply (ONLY) of Malonyl-CoA , then it may further depends on levels of other influencing factors like the activity of AMPK within that cells. AMPK as is activated by increase in AMP/ATP ratio, which is indication of fuel requirement or fasting or exercise also influence balancing or absorption of Malonyl Co-A.

Also Malonyl CoA as you may know is indication of plenty (sufficient energy). Therefore,its balance or exogenous source requirement/absortion within cell depends on another factor at the same time.

To check the effect of inhibition of pathways in animals/humans,you may go for profile of Urine organic acids and compare it with standard /normal.
If small animals then you may study effect by evaluating it with decrease in body temperature or weight loss etc.

Further, specify your queries & Wish you best of luck for your project !
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#3
sir, to be specific, i will be experimenting with trypanosomatids.so the problem of gram +ve/-ve will not arise.i want to inhibit the first step of FA synthesis where acetyl-CoA is converted to malonyl-Coa. So to check the effect, i want to supply malonyl-CoA and see if the phenotype is recovered to that of untreated cells.so, will this approach work?
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#4
(10-17-2012, 01:34 AM)billamin Wrote: sir, to be specific, i will be experimenting with trypanosomatids.so the problem of gram +ve/-ve will not arise.i want to inhibit the first step of FA synthesis where acetyl-CoA is converted to malonyl-Coa. So to check the effect, i want to supply malonyl-CoA and see if the phenotype is recovered to that of untreated cells.so, will this approach work?

Dear Billamin,
If possible for your project then try supplementing the culture media
with Oleate. It has the formula CH3(CH2)7CH=CH(CH2)7COOH.
It will work, good luck !
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#5
thanks for the reply sir. should i also suppliment oleic acid with palmitic acid (a combination of saturated & unsaturated FA precursors or the cell is able to take care of this matter using its own sets of saturases & desaturases)? and regarding my other reagent->Malate. A part of my experiment actually involves inhibiting the conversion of pyruvate to oxalocetate. Since oxaloacetate transporters are not present in the mitochondrial membrane, i have assumed that the same is also the case in the plasma membrane. Is this a valid assumption? Focusing on this assumption, i hypothesised that malate, whose transporters are present in mitochondrial membrane, and is readily converted to oxaloacetate by MDH, can be supplimented in place of oxaloacetate. please help me out in this matter. thank you.
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#6
(10-17-2012, 07:55 AM)billamin Wrote: thanks for the reply sir. should i also suppliment oleic acid with palmitic acid (a combination of saturated & unsaturated FA precursors or the cell is able to take care of this matter using its own sets of saturases & desaturases)? and regarding my other reagent->Malate. A part of my experiment actually involves inhibiting the conversion of pyruvate to oxalocetate. Since oxaloacetate transporters are not present in the mitochondrial membrane, i have assumed that the same is also the case in the plasma membrane. Is this a valid assumption? Focusing on this assumption, i hypothesised that malate, whose transporters are present in mitochondrial membrane, and is readily converted to oxaloacetate by MDH, can be supplimented in place of oxaloacetate. please help me out in this matter. thank you.

Dear Billamin, Add palmitic acid along with oleic acid as suppliment. As like mitochondrial membrane, Oxaloacetate transporters are also not present in cell membrane.
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Can cells be grown in exogenous supply of Malonyl-CoA and Malate? Need Help00