Lecture 23: Nitrogen Metabolism Flashcards

1
Q

proteasomal complex

A

releases oligopeptides

degrade these into individual amino acids

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2
Q

urea cycle

A

uses nitrogen from NH4+ and amino acid aspartate
generate urea
excrete to maintain daily nitrogen balance

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3
Q

what do we use the carbon skeleton of amino acids for?

A

energy converting rxns

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4
Q

what do we do with nitrogen after taking apart amino acids

A

remove it to avoid ammonia toxicity

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5
Q

what purpose does nitrogen fixation and assimilation serve?

A

FIXATION: in bacteria
- main way atmospheric nitrogen is made into ammnia and nitrogen oxides
ASSIMILATION: incorportate ammonium into amino acids (glutamate and glutamine)

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6
Q

Key enzymes

A

Bacterial Nitrogenase complex: redox rxns and ATP hydrolysis to make NH2 2NH3
Glutamine Synthetase: in all organisms NH4+ into glutamate to make glutamine
Glutamate synthetase: in bacteria and plants remake glutamate so we can continue glutamine synth
Glutamate dehydrogenase: all organisms. interconverts glutamate, NH4+ and alpha ketogluterate

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7
Q

Three processes that fix N2 into the biosphere

A

Rhizobium bacterium
Atmospheric Lightening
Haber Process

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8
Q

Haber process

A

important for agriculture
been around a long time
1 part nitrogen, 1 part H2 gas combined, some converted to ammonia gas
not very effcient, so recycle so that not used parts can be used

has to be very high temp and pressure

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9
Q

Nitrogen fixation in bacteria

A

nitrogenase protein required: 2 components
1 component binds to ATP and obtains electrons to reduce triple bond to double bond
(2nd component) nitrogenase component takes it from nitrogen to ammonia

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10
Q

how many ATP to convert 1 nitrogen to 2 ammonia???

A

16

2 ATP for every electron used by the nitrogenase protein

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11
Q

why so much ATP?

A

the bond NRG of N2 is 930 kJ/mol

this is not easy to split!

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12
Q

Dimine has two fates

A

can become hydrazine or go straight to NH3

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13
Q

Nitrogen assimilation in plants: HIGH NH4+ levels in soil

A

plants use glutamate dehydrogenase rxn to directly incoperate NH4+ into glutamate using ketogluterate

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14
Q

standard free energy charge is +30… so how do we get it to work??? (for nitrogen assimilation via glutamate dehydrogenase)

A

need high concs of NH4 in soil!!!!
(high substrate concs)

otherwise, we release reactants from glutamates (if levels not hight)

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15
Q

Nitrogen assimilation in plants: LOW NH4+ levels in soil

A

we need 2 enzymes/reactions:
glutamine synthetase
glutamate synthase: makes more glutamate that we can use for glutamine synth…provides more substrate to drive reaction forward

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16
Q

Net reaction of nitrogen assimilation: LOW NH4+ in soil

A

1) glutamate+NH4+ +ATP=glutamine
2) glutamine+alphaketoglutamate+NADPH= 2 glutamate

use ATP AND NADPH

net rxn: alphaketogluterate+ NH4++ ATP + NADPH=glutamate

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17
Q

whats the difference between low and high (besides needed 2 vs 1 enzymes)

A

low use ATP and NADPH

high uses just NADPH

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18
Q

assimaltion of nitrogen into amino acids done by…

A

aminotransferase enzymes

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19
Q

aspartate aminotransferase

A
uses PLP (vitamin B6) as a coenzyme to help with amino group transfer
nitrogen temp attached at PLP

glutamate combined with the aminotransferase
PLP attaches to nitrogen temporarily
remove nitrogen from glutamate=alphaketogluarate
nitrogen transferred to OAA to from aspartate

20
Q

What pathway provides the OAA needed for the aspartate aminotransferase reaction reaction?

A

citrate cycle

21
Q

amonia cycle

A

NH4 in soil comes from decomposition, soil bacteria, fertilizers

converted to NO2- and NO3- by soil bacteria (nitrite and nitrate)

plant roots absorb this and make it NH4+

22
Q

Positive nitrogen balance

A

young children and pregnant women

they accumulate nitrogen to support protein synth

23
Q

negative nitrogen balance

A

seen in starvation, elevated rates of protein break down

24
Q

Digestion of dietary proteins

A

proteases secreted: they secrete inactive pepsinogen
pepsinogen becomes pepsin, which breaks down the dietary protein (stomach)
more proteases in small intestine continue to break down

end result: amino acids and small peptides

25
What do proteases do
break down proteins into amino acids and peptides
26
why do we first have pepsinogen: which is inactive?
so its active only in stomach (low pH) | so that it doesnt break down proteins in cells
27
where are proteases made? How are they activated
pancreas | when they reach low pH of stomach, another protease cleaves them and activates them
28
degradation of cellular proteins. | why?
to regulate enzyme function | if you degrade the enzyme... its gone and you can't use it
29
how to break down proteins in cells
in lysosomes proteosome complexes left with amino acids at the end
30
breakdown of proteins in cells: lysosomes
phospholipid bilayer | low pH inside where enzymes that break things down are
31
breakdown of proteins in cells: proteosome complexes
ID proteins that have been modified with protein ubiquiton (ubiquitinated proteins) (tag for degredation) unfold proteins and break them into small peptides
32
What do we do with amino acids?
use whole thing again to make more proteins split it up - carbon skeleton to make pyruvate, OAA, acetyl CoA for citrate cycle - nitrogen group: toxic, use it to make biomoelcs or excrete it
33
2 types of amino acids
glucogenic | ketogenic
34
glutogenic amino acids
form citrate intermediates or pyruvate when broken down
35
ketogenic amino acids
make acetyl Co-A or acetoacetyl CoA when broken down
36
Urea Cycle
to make soluble and non toxic urea from ammonium ions, bicarbonate, aspartate ammonium ion either comes down from breakdown of amino acids or from bicarb coms from citrate cycle aspartate comes from glutamate AMINO ACIDS
37
what is accomplished by the urea cycle
removal of excess nitrogen from body synthed in liver exported to kidneys leaves through baldder
38
regulated enzymes of urea cycle?
carbamoyl phosphate synthase 1 (first step in pathway)
39
where does reaction happen?
first step happens in mitochondrial matrix | exportation into cytosol
40
urea cycle
review slide 22
41
aspartate-arginiosuccinate shunt
link between urea and citrate cycles | provides substrates so they can continue
42
Where does amino acid-derived nitrogen enter the urea cycle?
at the glutamate and NH4 (see slide 23)
43
review slide 23
review slide 23
44
Urea Cycle disorders: Arginosuccinase deficiency
you can't convert argininosuccinate so you're on low protein diet except you give them lots of arginine!!! because they can't make it
45
would suplementing with ornithine also work?
in theory yea, it still helps us to make citrulline; which is the goal but we don't do it b/c its not as practical: arginine is really availible and is easily taken up