Cellular Regulation of

Anabolism and Catabolism

Erich Roth
Medizinische Universität Wien
Klinik für Chirurgie/Forschungslaboratorien
 Cellular Regulation of Anabolism
and Catabolism

• Amino acid metabolism

• Protein synthesis
• Proteindegradation
• Energy metabolism

• Hormons, Interleukins, growth factors

• Signaling molecules
• Neuromuscular disorder
Composition of human
muscle

g/kg
Dry solid 230
Extracellular water 120
Intracellular water 650
 Three ways to atrophy of skeletal
muscle

• Protein catabolism mediated by catabolic

factors
• Disuse of skeletal muscle
• Sarcopenia of old age
 One-Year Outcomes in Survivors of the
Acute Respiratory Distress Syndrome
Herridge MS et al. New England J Med 348 (2003)

• …the patients have persistent functional

limitation one year after discharged from
the ICU largely as a result of muscle
wasting and weakness
• ..the impaired muscle function had an
important effect on the long term
outcomes on these patients
Disuse: Immobilisation of the knee
– effect on thight muscles

Four weeks:

loss of 53 % of muscle strength of knee

extension
Quadriceps circumference was decreased
for 21 %
 Sarcopenia of Old Age
• Reduction of muscle mass:
10 % until the age of 60 yr
30 to 40 % until the age of 80 yr
up to 30 % between 80 to 90 yr

• Reduction of muscle strength

> 60yr
3 % loss of grip stength per year: male
4 % loss female
 Strength, but not muscle mass, is
associated with mortality in older adults.
AB. Newman et al. J Gerontology 61A;2006

Men, leg strength, and mortality.
Kaplan–Meier survival curves for leg strength groups (<90, 90–
<130, 130–<170,  170 Nm). Intervals of 40 Nm of quadriceps
strength were used to approximate men's standard deviation =
33.8 and to distribute the number of events
Men, grip strength, and mortality.
Kaplan–Meier survival curves for grip strength groups (<30, 30–
<40, 40–<50, 50 kg). Intervals of 10 kg of grip strength were
used to approximate men's standard deviation = 8.5 and to
distribute the number of events
Muscle mass, muscle strength, and muscle fat
infiltration as predictors of incident mobility
limitations in well-functioning older persons

M Visser et al, J Gerontol A Biol Sci Med Sci (2005)

Muscle fiber specific apoptosis and TNF-alpha signaling in
sarcopenia are attenuated by life long calorie restriction
Phillips T et al. FASEB 19 (2005)
Muscle fiber specific apoptosis and TNF-alpha signaling in
sarcopenia are attenuated by life long calorie restriction
Phillips T et al. FASEB 19 (2005)
 Sarcopenia: An inflammatory
process with an increased insulin
resistance?
Regulation of atrophy
 Catabolic stimuli in critically ill
patients: supposed causes

• Increased catabolic/Inflammatory
mediators
• Reduced anabolic stimulators

• Polyneuropathy
• Loss of electrical excitability of the
sarcolemma
• Neuromuscluar blockade by drugs
Catabolic factors of skeletal muscle
catabolism
• Interleukins: TNF-α, IL-1, IL-6, IFN-γ
• Myostatin: member of transforming
growth factor family (increased in AIDS)
• Glucocorticoids (affecting the IGF-I
pathway)

• Reduced levels of growth hormone

• testosteron
 Denervation
• Denervated muscle quickly atrophy due to
increased proteolysis
• Long-term denervation leads to myofibre
death
• Denervation as a result from blocked
signals of physical disruption at the
neuromuscular junction (synapse)
 Signalling

mechanischer Reiz IGF-1

_
Apoptosis
IRS-1 (BAD / BAX)
Integrine/Vinculin/Talin RAS PI3K
SHIP2
PI3K PTEN eIF-2B
RAF Glucocorticoide GSK3ß Translation
Akt-1 initiator
Cain Akt-1
MCIP1 Ca+-Calmodulin
+ Rapamycin
MEK
_
+ FKBP12
FOXO mTOR
mTOR
+ amino acids
Calcineurin
NFAT ERK

PHAS1 = 4E-BP1
P70S6K
Hypertrophy Translation-initiator
MGF Proliferation + elongation
eIF-4E
MAFbx Translation-initiator
Nucleus MURF1
 DNA Genome
Transkription

RNA
Translation

Protein Proteome

Metabolic
Metabolome
action
 Gene regulation

Transcriptional profile of a myotube

starvation model of atrophy.
E.J.Stevenson et al. J Appl Physiology 98;2005
 Altered Marker genes

• Muscle contraction (4)

• Structural components (6)
• Cytoskeleton (4)

• Proteolytic enzyms (3)

• Biosynthesis (3)

• Oxidative stress (1)

• Signaling (15)
• Growth factors (9)
 Protein level

Protein synthesis
Protein degradation
Protein synthesis
Stress and protein turnover

 Slow wasting condition

is found in mild injury,
mal-nutrition, cancer,
immobilization.

 Rapid wasting occurs

after severe injury,
burns and infection.  
 One genome - different
proteomes

• > 35,000 coding genes in higher organisms

• Differently spliced
• Posttranslational modified
• 100,000 proteins / organism
• Selectively expressed in tissues and cells
• > 20,000 proteins / cell
 Proteomic analysis of rat soleus muscle
undergoing hindlimb suspension-induced atrophy

RJ Isfort et al, Proteomics 2 (2002)

Proteomic analysis of altered protein expression in
skeletal muscle of rats in a hypermetabolic state
induced by burn sepsis

X Duan et al, Biochem J (2006)

Significantly regulated proteins in EDL muscle
following burn-CLP treatment

X Duan et al, Biochem J (2006)

 Proteomics

Energy Occurrence of Stimulation of

regulation oxygen radicals Protein degradation
Protein degradation
Muscle protein breakdown and the critical role of
the ubiquitin-proteasome pathway in normal and
disease states
Abnormal proteins Ubiquitin-
Proteasome
pathway
Short-lived normal proteins

Long-lived normal proteins

Proteins of the Lysosomes

Endoplasmic Reticulum

Extracellular proteins
Surface Receptors
Mitochondrial
Mitochondrial proteins proteases

SH Lecker et al, J Nutr (1999)

 Disuse
• Calcium overload
• Increased calpain activity, increased
degradation of sarcomeric proteins
• Stimulation of calpases activities (ROS?)
• Degradation of intact actomyosin
complexes
• Proteasome system degrade monomeric
contractile proteins
 Proteasome mediated proteolysis
• Either by 20S or 26S proteasome
• 26S is composed of the 20S core proteasome with a
regulatory 19 S complex connected to each end
• 19S complex posses ATPase activity

• 26S pathway – ubiquitin convalently binds to protein

substrates
• Unfolding of protein by 19S ATP dependent process

• Degradation of the protein in the 20S core proteasome

(oxidized protein degradated without ubiquitination)
 ..moreover
• Ubiquitin-activating enzymes (E1)
• Ubiquitin-conjugating enzymes (E2) Ubiquitin
protein ligase enzymes (E3)

• Specific ligases: atrogin 1

muscle ring finger-1
(both upregulated by ROS) out of stimulation of two
genes: MAFbx (muscle RING Finger !)
MuRF1 (muscle Atrophy F-box)
(upregulated 10 times by IL-1 and Dexamthason)
B Biedermann, Schweiz Med Forum 11 (2002)
Metabolism

Energy metabolism
Oxygen Radicals
 Energy metabolism in sepsis
• Decrease in oxygen extraction – increase
in tissue oxygen tension
• Reduced levels of ATP and ADP
• Increased level of AMP – AMPK activation
• Mitochondrial dysfunction through an
impairment in complex I mediated
respiration
• 30 % reduction in the ability to utilize
oxygen during exercise
 Consequences of AMPK activation on
metabolism during single bouts of exercise

WW Winder, J Appl Physiol. 2001; 91:1017-28

 Decreased antioxidative
metabolites in skeletal muscle

• Chemoluminescensce + 100 %
• Mn-Superoxiddismutase - 46 %
• Catalase - 83 %
• Glutathioneperoxidase - 55 %

S.Llesuy et al. Free Rad Biol Med 16 (1994)

Amino acid Metabolism
 Muscle Glutamine

control

Starvation (4 days)
Abdominal Sepsis
Injury & Infection

(Survivors)
Postop.
20

Abdominal Sepsis
(Nonsurvivors)
mmol/l

10

0
 Roth E, et al.: Clin Nutr 1:25, 1982
 Vinnars E, et al.: Ann Surg 182:665, 1975
 Askanazi J, et al.: Ann Surg 192:78, 1980
 Elwyn DH, et al.: in Walser M & Williamson JR (eds.) Metabolism and Clinical Implications
of Branched Chain Amino Acids and Ketoacids. Elsevier NY 1981, pp 547-
552
 AMPK pathway
catabolic pathways ↑
ATP↓ AMPK anabolic pathways ↓
protein synthesis ↓
GLUTAMINE ? Cellular redox state TRX:ASK1 ↓ JNK
DEPLETION GSH↓ redox-sensitive
kinases NF-kB
AP-1
Osmo-signaling
Fas ↑
delayed Hsp70-induction apoptosis
cell volume ↓
Erk ↓
p38 ↓ autophagic-proteolysis ↑

Translation p70s6k
mTOR ↓ 4E-BP1 translation ↓
...

glutaminyl-tRNA synthetase
QRS:Gln:ASK1 complex ↓ JNK
Anabolic Regulation
 Muscle anabolism
• Increased IGF-I, especially of one isoform
stimulated especially by strechting
exercise
• Growth hormone
• Testosterone
• IL-12, IL-15
• Overexpression of the oncogene „ski“
 Growth factors, especially IGF-I,
affect neuronal function

• Myelination
• Prevention of apoptotic death
• Stimulation of axonal sprouting
• Repair of damaged axons
 Any hope ?
Which concepts are reliable?
 Electrostimulation
• Fast-to-slow muscle conversion

• Increased respiratory chain activity and

efficiency

• Increased amount of cytochrome a, a3,

b562, c, c1
STRETCHING
G.Goldspink et al. J.Physiol.1999:516
 IL-6

TNF

IL-6
The
anti-inflammatory
effect of exercise
AM Petersen et al, J Appl Physiol (2005)
Metabolic effects of IL-6 released by
muscle fibers

IL-6

IL-6

IL-6

AM Petersen et al, J Appl Physiol (2005)

Geoff Goldspink
Royal Free & University College Medical
School, London, UK

MGF and the regulation of

muscle strength.
Alternative Splicing of the Human
IGF-I Gene
3 4 6

IGF – IEa (systemic)

Hormones 3 4 5
effect on the
liver
IGF - IEb
1 2 3 4 5 6
Promoter 1 Promoter 2

Human IGF–I gene

3 4 5 6
Mechanical
signals including
local cell damage MGF (autocrine)
reading frame *
shift 49 base insert
The effect of treating the muscles of mdx dystrophic mice with MGF
in a plasmid vector after only 3 weeks on muscle strength

40 ***
force compared to their contralateral
Percentage of treated muscle tetanic

35
30
untreated muscle (%)

25
20
15
10
5
0
-5
-10

MGF Vector
 MGF peptide use for generic
treatment for damage and repair
1) Rescue and repair of muscle in muscular
dystrophy, (ALS) and other diseases.
2) Postural problems arising from muscle
weakness.
3) Muscle cachexia in cancer, HIV, COPD,
cardiac and renal disease.
4) Age-related muscle loss – sarcopenia.
 Nutrition
• Can reduce but not avoid protein
catabolism

• Exert anabolic effect

Short-term bed rest impairs amino acid-
induced protein anabolism in humans

• 14 day period of strict bed rest or controlled

ambulation
• Weight-maintaining diet

• 3h infusion of an amino acid mixture

• Determination of whole-body protein kinetics

 Bed rest leads to reduced stimulation of protein

synthesis by amino acid administration
G Biolo et al, J Physiol (2004)
 Latency and duration of stimulation of
human muscle protein synthesis during
continuous infusion of amino acids
J Bohé et al, J Physiol (2001)

Time course of Time course of rate of synthesis of

serum insulin and glucose mixed muscle proteins
 Rates of synthesis
of mixed muscle proteins and muscle fractions
(myofibrillar, sarcoplasmic and mitochondrial proteins)

J Bohé et al, J Physiol (2001)

 Protein synthesis during and post-
exercise

• Protein synthesis is decreased during exercise

• PS is increased post exercise untill hr 38

• Exercise stimulates protein translation by

increasing the group 4 eurkaryotic initiation
factor eIF4E
• Post meal exercise composition (CH+
proteins/AAs) influence the availability of eIF4E
 Pathophysiology: Impressive for
me…
• Prognostic importance of muscle strength
and fat-infiltration
• Role of oxygen radicals in protein
degradation– antioxidants
• Caloric restriction: reduced degradation
process – insulin resistance ??
• Role of IL- 6: inflammatory vs anabolic
stimulus?
 Conclusion
• Pathogenesis of protein catabolism is
multifactorial…..

• Therapeutical interventions:
• Exercise – electrostimulation (?)
• Appropriate nutrition
• Endocrine and mediator-directed strategies
• Behavior, emotional interactions – well-being,
anti-stressing
Muscle Fibre Size After Injection Of MGF
cDNA
1200
25% increase
in x sectional
1000 area

800
number of fibres

injection
600 control

400

200

0
up to 800 801-2000 2001-3000 3001-4000 4001-5000 5001-6000

area of fibres (square micrometer)

 Molecular events after stretching
• Increased myoblast proliferation
• Increased COX2 mRNA stimulation of
mTOR and rapamycin-p70 S6 kinase
• Satellite cell activation by HGF and NO
• Increase of mechano growth factor (MGF)
 Coupling of Voltage-sensitive Sodium
Channel Activity to Stretch-induced Amino
Acid Transport in Skeletal Muscle in Vitro

• Stretching of tissue-cultured skeletal

myotobes stimulates amino acid uptake

• Serum factors are not required for stretch-

induced AA uptake

• Alterations in the cell‘s voltage-sensitive

sodium channel and sodium pump activity

HH Vandenburgh, S Kaufman, J Biol Chem (1982)

Ubiquitin – Proteasom System

Mitch et al., New England Journal of Medicine 1996

 Disuse
Spinal cord isolation:

• Atrophy of slow and fast extensor muscles

• The slow rat soleus atrophied by ~ 50 % within 15 days
• Myofibrillar protein content, myosin heavy chain ~ 50 %
• Actin maintained at control level

Molecular mechanism:
• reduction in ribosomal RNA and protein translational
capacity
• Unsufficient RNA substrate for translating key
-sarcomeric proteins compromising the myofibril fraction
 Disuse of Skeletal Muscle

Reduced muscle tension

Disuse
(e.g. bedrest, immobilization, denervation,
Conditions:
unloading, spaceflight)

• Akt • Glucocorticoids
Triggers & • mTOR • myostatin
Signals: • p70S6kinase • NF-kappaB
• 4E-BP1 • Reactive oxygen species

or

Target Protein synthesis Protein degradation

Systems: rate rate

RW Jackman, SC Kandarian. Am J Physiol Cell Physiol (2004)

Cell volume and hormone action

TIPS-October 1992 [Vol.13]

Current awareness
 Muscle tissue changes with aging
• Reduction in muscle cell number, in
muscle twitch time and force
• Reduction in sarcoplasmic reticulum
• Reduction in calcium pumping capacity
• Disorganisation of sarcomere spacing
• Centralisation of muscle nuclei along the
muscle fiber
Muscle Cachexia: Current Concepts of Intracellular
mechanism and Molecular Regulation
PO Hasselgren, JE Fischer Ann Surg 233 (2001)