Humboldt State University ® Department of Chemistry

Richard A. Paselk

Chem 438

Introductory Biochemistry

Spring 2007

Lecture Notes: 16 April

© R. Paselk 2006
 
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Fat Metabolism 3

KETONE BODIES

Introductory discusion of fat metabolism, exercise, and fasting.

Fatty acids can be used as the major fuel for tissues such as muscle, but they cannot cross the blood-brain barrier, and thus cannot be used by the central nervous system (CNS). This becomes a major problem during starvation (fasting), particularly for organisms such as ourselves in which CNS metabolism constitute a major portion of the resting basal metabolic rate. These organism must provide glucose to the CNS to provide for metabolic needs, and thus during the initial fasting period must break down substantial amounts of muscle tissue (protein) to provide the amino acid precursors of gluconeogenesis. Obviously the organism could not survive long under such a regime. What is needed is an alternate fuel source based on fat rather than muscle. The so-called ketone bodies serve this function:

Note that only two of the ketone bodies are in fact ketones, and that acetone is an "unintentional" breakdown product resulting from the instability of acetoacetate at body temperature. Acetone is not available as fuel to any significant extent, and is thus a waste product.

CNS tissues can use ketone bodies any time, the problem is the normally very low concentrations (< 0.3 mM) compared to glucose (about 4 mM). Since the KM's for both are similar, the CNS doesn't begin to use ketone bodies in preference to glucose until their concentration exceed's the concentration of glucose in the serum.

The system becomes saturated at about 7 mM. The limiting factor in using ketone bodies then becomes the ability of the liver to synthesis them, which requires the induction of the enzymes required for acetoacetate biosynthesis. Normal glucose concentrations inhibit ketone body synthesis, thus the ketone bodies will only begin to be synthesized in high concentrations as serum glucose concentrations fall. As an example, ketone bodies might start at about 0.1 mM after an overnight fast, rise to 3 mM after a 3 day fast, and go to 7-8 mM with prolonged fasting (>24 days).

Ketogenesis

Ketogenesis occurs in the matrix of liver mitochondria. Fatty acids are first broken down to acetyl CoA via beta-oxidation (providing energy for liver metabolism from the reducing equivalents generated). The acetyl CoA is then used in ketogenesis:

Depending on the status of the liver, acetoacetate can now be reduced to give D-beta-hydroxybutyrate, which delivers more reducing equivalents, and thus ATP equivalents, to the peripheral tissues at the expense of the liver:

Ketone Bodies as Fuel

The ketone bodies are water soluble and are transported across the inner mitochondrial membrane as well as across the blood-brain barrier and cell membranes. Thus they can be used as a fuel source by a variety of tissues including the CNS. They are preferred substrates for aerobic muscle and heart, thus sparing glucose when they are available.

In the peripheral tissues the ketones must be reconverted to acetyl CoA in the mitochondria:

The energy provided to the peripheral tissues from acetoacetate and for beta-hydroxybutyrate are shown below:
Reaction Energy Product Factor Multiplier ATP Equiv.
CoA transferase succinate (-GTP) 1 1 -1
Kreb's NAD+ DH's NADH 2.5 x 3 2 15
Kreb's GTP 1 2 2
Kreb's FADH DH FADH2 1.5 2 3

Acetoacetate Total:

19
Butyrate DH NADH 2.5 1 2.5

Butyrate Total:

 21.5

Note the P/O ratios for the ketone bodies: Acetoacetate = 19 ATP / 8 O = 2.38; Butyrate = 21.5 ATP / 9 O = 2.39 which are higher than we calculated for palmitate (2.3), but again lower than for glucose (2.67).

These reactions can be thought of as giving the liver overall control of fat metabolism. Lower vertebrates store fat in the liver. In a sense adipose tissue can be thought of then as "extended liver" tissue metabolically. Note that the liver can adjust the amount of reducing equivalents, and thus ATP equivalents, it sends to the peripheral tissues by adjusting the amounts of acetoacetate vs. beta-hydroxybutyrate it exports. Thus the percentage of the free energy distributed between the tissues is shown below:

 Compound \ Tissue Liver Peripheral Tissues
3-hydroxybutyrate 17% 83%
Acetoacetate 26% 74%

Fatty Acid Biosynthesis

[Reference: Wakil, S. J., Stoops, J. K., and Joshi, V. C., Ann. Rev. Biochem. 52, 537-79 (1983).]

Chemically fatty acid biosynthesis is a review of of beta-oxidation and pyruvate carboxylation. Want to reverse the reactions of beta-oxidation. But beta-oxidation is highly favorable as is, and want synthesis to also be favorable - obviously some variations in the pathways are needed.

Two reactions enable the synthesis pathway:

The first step in fatty acid biosynthesis is to activate acetyl-CoA by the addition of a carbon dioxide using Acetyl-CoA carboxylase. This reaction is chemically identical to the Pyruvate carboxylase reaction seen earlier (8 April 2003, Gluconeogenesis):

This reaction is physiologically irreversible and is the flux generating or first committed step of fatty acid biosynthesis. As expected it is regulated. In mammals acetyl-CoA carboxylase is a large enzyme existing as inactive protomers (560,000 MW, 4 subunits, one biotin), which can assemble into active filaments (4 - 10 million MW).

Formation of the filaments (activation) is

As we shall see these are excellent indicators of the fuel status of the cell.

In addition to the activator/inhibitor controls of citrate and fatty acyl-CoA, the enzyme is also under hormonal control (note the similarities to glycogen control):

In mammals Fatty Acid Synthase (FAS) [overhead] catalyzes fatty acid synthesis on a homodimeric enzyme, each monomer of which has seven catalytic activities, and eight sites! (In bacteria such as E. coli there are seven separate enzymes plus an acyl-carrier protein. Plants also have individual proteins for the various activities which are associated in a quaternary complex. In eukaryotes other than plants the FAS are complexes of multifunctional proteins. Note: the numbers in the sites correspond to the numbered reactions of the FA Synthase reactions diagram, below.) The enzyme weighs approximately 500,000 Daltons. [Reference: Maier Timm, Simon Jenni and Nenad Ban. "Architecture of Mammalian Fatty Acid Synthase at 4.5 A Resolution." Science 311(3 March 2006) 1258; mini review comparing fungal and mammalian enzymes - Smith, Stuart. "Architectural Options for a Fatty Acid Synthase." Science 311(3 March 2006) 1251.]

There are two carriers on this complex.

Looking at the the Fatty Acid Synthase reactions (packet or Figure 16.4 & 5):
We now have a beta-ketoacyl group ready to go through the reverse of the reactions of beta-oxidation.


Pathway Diagrams

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Last modified 16 April 2007