Biology 301: Biochemistry Coenzymes ICoenzyme A, Thiamine Diphosphate, Pyridine Nucleotides, Flavins, and Lipoic Acid

I.

Coenzymea small organic molecule bound at least transiently to the active site of an enzyme by covalent or noncovalent bonds, which participates in catalysis Coenzyme A
A.
NH2 N N N O N O O P OOPO32O O P OO N CH3 OH H N H CH3 O O SH

II.

Structure
Pantothenic acid, vitamin B5 5-10 mg d-1

OH

Coenzyme A

B.

Function 1. Activation of acyl groups

O R

toward transfer by nucleophilic displacement

O Y- + R SCoA R Y O +
-SCoA

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BIOLOGY 301: BIOCHEMISTRY LECTURE 11: COENZYMES ICOENZYME A, ETC. 2. Activation of a hydrogen atom adjacent to the carbonyl of a thioester
O H C H BSCoA H C SCoA OH CSCoA O

III.

Thiamine pyrophosphate
A. Structure
Thiamine, vitamin B1 1.2-1.5 mg d-1 NH2 N H3C N N+ H3C S O O P OThiamine pyrophosphate O O P OO-

1.

A deficiency of thiamine in the diet results in beriberi, characterized by numbness, burning sensations, muscle wastage, heart failure Seen only in people with severe dietary restriction

2.

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BIOLOGY 301: BIOCHEMISTRY LECTURE 11: COENZYMES ICOENZYME A, ETC. B. Function: -cleavage and -condensation, e.g. pyruvate decarboxylase
S H NH2 N H3C N H3C N+ CH3 N N R S
-

NH3+ N+

CH3

Reasonably stable ylid H+ O H3C Pyruvate COOH S

-

O HO

OS

H+ HO

S N R'

CH3 N+ R' CH3

N+ R'

CH3 CO2

CH3

CH3

O Enz-BH + H3C H +
-

S N+ R'

R Enz-BCH3 H O

S N+ R'

CH3

CH3

Acetaldehyde

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BIOLOGY 301: BIOCHEMISTRY LECTURE 11: COENZYMES ICOENZYME A, ETC.

IV.

Pyridine nucleotide coenzymes

A.
NH2

Structure of nicotinamide adenine dinucleotide (NAD+)

O N O O O P ONAD+ OH O O N+ N O O OH OH P ONH2 Nicotinamide (niacin), vitamin B3 16-20 mg d-1

N N

2'

OH

Esterified to phosphoric acid in NADP+

1.

A deficiency of niacin in the diet results in pellagra, characterized by dermatitis, diarrhea, and dementia. Seen in cultures where corn (maize) is the chief source, and the corn is not treated with alkali before consumption.

2.

B.

Function 1. A hydride acceptor in 2-electron redox reactions

HN O O O P ONAD+ OH O O N+ N O O OH OH P OO H H O NH2 N R OH NADH NH2

NH2 N N

2'

2.

NAD+ tends to be used for catabolic reactions, while NADPH tends to be used for anabolic reactions.

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BIOLOGY 301: BIOCHEMISTRY LECTURE 11: COENZYMES ICOENZYME A, ETC. 3. Different enzymes seem to be specific for different prochiral hydrogens
pro-R pro-S O H H NH2 N+ R

4.
N+

Glyceraldehyde-3-phosphate dehydrogenase
OH H O OH OPO32Glyceraldehyde-3phosphate H N OH OPO32NADH HOPO32NH2 H O OH OPO32NH2
-S-Enz

N+

H H

S-Enz OH

O NH2

OPO32-

O HO P OO-

S-Enz

S-Enz

O
-S-Enz

HO

P O-

O OH OPO32-

1,3-Bisphosphoglycerate

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BIOLOGY 301: BIOCHEMISTRY LECTURE 11: COENZYMES ICOENZYME A, ETC.

V.

Flavins
A. Structure
Adenosine monophosphate (AMP)

Riboflavin-5'-phosphate (FMN) N O O OH OH OH Riboflavin, vitamin B2 1.3-1.7 mg d-1 H H3C N H N N H3C N O H O OH OH P OO O P OO O N

NH2 N N

Flavin adenine dinucleotide

B.

Function 1. 2. Usually tightly bound to enzymes A hydride acceptor in 2-electron redox reactions FAD + RH2 3. FADH2 + R

FAD is a stronger oxidant than NAD+ Reaction Riboflavin + 2 H+ + 2 eNAD+ + H+ + 2 eDihydroriboflavin E (V) -0.208 -0.32

NADH

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BIOLOGY 301: BIOCHEMISTRY LECTURE 11: COENZYMES ICOENZYME A, ETC. 4.

FAD H3C

Thus FAD can oxidize NADH:

R N HB-Enz N N O FADH2 H3C R N H N N H H O R' H3C H N H R' O H
-B-Enz

H3C H

N NADH R NAD+

N+ R

5.

Flavins can also mediate one-electron transfers

Hydroquinone form R N N O H3C N H N N H O H3C N H O H O

Quinone form R H3C N

H3C

H (one electron + one proton) H3C

R N N N H3C N H O H O H (one electron + one proton)

Semiquinone form

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BIOLOGY 301: BIOCHEMISTRY LECTURE 11: COENZYMES ICOENZYME A, ETC.

VI.

Lipoic acid
A. Structure
COOH S S Lipoic acid

1.

Although lipoic acid has to be supplied in the culture medium for Tetrahymena geleii, no nutritional deficiency has ever been documented in humans. Lipoic acid is in mammalian tissue in extremely low quantities.1

2. B.
H+ O H3C

Function: an acyl carrier, e.g. in pyruvate dehydrogenase2

R'' Pyruvate COOH S
-

O HO

OS

H+

S HO

S S N R'

CH3 N+ R' CH3

N+ R'

CH3 CO2

CH3

CH3

R'' H Enz-BH + CoASH S O S + CH3

-

R'' S N+ R' R Enz-BCH3 H H S O CH3 N+ R' CH3 S S R

Lipoic acid was first purified and characterized by Lester Reed of the University of Texas and a team from Eli Lilly and Company. In 1949 they isolated 30 mg of lipoic acid from 10 tons of water-soluble liver extract. 2 The eukaryotic pyruvate dehydrogenase complex is the largest multienzyme complex known. It consists of about 22 molecules of pyruvate dehydrogenase, about 6 molecules of dihydrolipoamide dehydrogenase, and a 60-subunit dihydrolipoamide acetyltransferase core. See ZHOU, Z. H., D. B. MCCARTHY, C. M. O'CONNOR, L. J. REED, & J. K. STOOPS. The remarkable structural and functional organization of the eukaryotic pyruvate dehydrogenase complexes. Proc.Natl.Acad.Sci.USA 98(26): 1480214807, December 18, 2001.

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BIOLOGY 301: BIOCHEMISTRY LECTURE 11: COENZYMES ICOENZYME A, ETC.

R'' H S O CH3 S H+
-SCoA

Acetyl CoA O + H3C NADH SCoA H FAD S S H R''

NAD+

FADH2 R'' S S

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BIOLOGY 301: BIOCHEMISTRY LECTURE 11: COENZYMES ICOENZYME A, ETC.

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