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Regula'on of mTOR by Growth Factors, Nutri'on and Exercise
Impact on Muscle Protein Synthesis

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Regula'on of mTOR by Growth

Factors, Nutri'on and Exercise

Impact on Muscle Protein Synthesis
John L. Ivy, PhD
University of Texas at Aus'n

mTOR
mTOR: Mammalian Target of Rapamycin
mTOR protein is a serine-threonine kinase
that belongs to the phosphoinosi<de 3-kinase
family of proteins
mTOR is found in two dis<nct protein
complexes
mTOC1 (rapamycin sensi<ve) cell growth
mTOC2 (rapamycin insensi<ve) cytoskeletal
organiza<on

mTOC1 Protein Complex

R
FKBP12

mTOR
Deptor
Raptor

mLST8
PRAS40

mTOR: mammalian target of rapamycin

Raptor: regulatory-associated protein of mTOR
mLST8: mammalian lethal with Sec 13 protein
PRAS40: proline-rich Akt substrate 40 kD
Deptor: DEP domain-containing mTOR-interac<ng protein
FKBP12: FK506 binding protein of 12 kD (R: rapamycin)

Protein Synthesis
The process by which individual amino acids
are connected by pep<de bonds in a specic
order dictated by the nucleo<de sequence in
DNA, which involves the processes of
transcrip<on and transla<on
Transcrip<on - is the rst step of gene
expression, in which a par<cular segment of DNA is
copied into RNA by the enzyme RNA polymerase
Transla<on - mRNA is decoded by a ribosome to
produce a specic amino acid chain or polypep<de

Protein Synthesis and Degrada'on

Hormone

DNA

Scarcolema
Hormone
Receptor
RNA
Polymerase

Promoter
TE

RE
Response
Element
Transcrip<on
Element

1. Transcrip'on

mRNA

Nucleus
TE

New
Pep<de
mRNA

3. Degrada'on

Muscle
Contrac'on

RA
Poly

Insulin

2. Transla'on
Amino Acids

Post Exercise
Nutri'on

Steps in Transla'on
1. Ini'a'on: The ribosome assembles
around the target mRNA. The rst
tRNA is adached at the start codon.
(ini)a)on thought to be rate-limi)ng
step for protein synthesis)
2. Elonga'on: The tRNA transfers an
amino acid to the tRNA corresponding
to the next codon. The ribosome then
moves to the next mRNA codon to
con<nue the process, crea<ng an
amino acid chain.

3. Termina'on: When a stop codon is
reached, the ribosome releases the
polypep<de.

mTOR Pathway
GS

mTOR

GSK3 P GS

eIF2

GDP

GSK3
4E-BP1

4E-BP1

eIF4E
p70S6K

P
eIF4E

eIF4E

eIF4F

eIF4A
eIF4G

p70S6K

eIF4B

rpS6

eIF4B

60S
Ribosome

Elonga'on: Pep'de Chain

eIF2B

rpS6

eIF2

GTP

40S

Ribosome

Transla'on Ini'a'on
Protein Synthesis
tRNAmet
GTP
eIF2

mTOR Ac<vity Required for

Increase in Muscle Mass
mTOR ac<vity is increased following resistance
exercise
Rapamycin inhibits mTOR ac<vty
Rapamycin administra<on during a resistance
training program blocks protein synthesis and muscle
hypertrophy
Rapamycin also blocks protein/amino acid ac<vated
protein synthesis
Rapamycin does not aect basal protein synthesis

mTOR Ac<va<on by Growth Factors

I R

IRS

Insulin/IGF-1
PI3K

Ptdlns(4,5)P2
P

Ptdlns(3,4,5)P3

Tuberous Sclerosis Complex

GTPase Ac<vator Protein (GAP)
Akt Inac<vates TSC

TSC2
TSC1

Ras Homolog Enriched in Brain

Ac<vates Cataly<c Subunit of mTOR

Akt
GDP

GTP
Rheb

Rheb

ADP

ATP

P
p70S6K

Phosphoryla<on of PRAS40
Ac<vates Raptor
Raptor Sequesters Substrate for
mTOR Cataly<c Subunit

CD Cataly<c Domain

mTOR

CD

Deptor
mLST8

Raptor
PRAS40
P

Ac<va<on of mTOR by Amino Acids

Amino Acid Pool
(L-Leucine)

Lysosome
Lysosomal Membrane

TSC2
TSC1

GTP

Insulin

Rheb

p70S6K

ADP

Akt

Rag A/B
Rag C/D

mTOR

CD

GTP

Ragulator

Deptor

ATP

Raptor

mLST8
PRAS40

GDP
Rag A/B

Gator

Rag C/D

Ragulator: Guanine nucleo<de exchange factor (GEF), when ac<vated will

convert GDP to GTP
Gator: GAP ac<vity towards Rags, will ac<vate GTPase and convert GTP to GDP
Rag: Ras related GTP binding , when GTP-bound it will recruit mTOR to
lysomomal membrane where it will bind and be ac<vated by Rheb
*Insulin ac<vates mTOR by a separate mechanism than AA and therefore their
eects on mTOR and protein synthesis are addi<ve
*Remove L-Leucine from lysosome stops ac<va<on of mTOR

mTOR Ac<va<on by Muscle Contrac<on

Insulin
I R

IRS

PI3K

IGF-1
Ptdlns(4,5)P2
Phospha'dylcholine

Ptdlns(3,4,5)P3

ERK 1/2
P
P TSC2

Phospholipase D

TSC1

Akt
GDP

GTP
Rheb

Rheb

ADP

Phospa'dic Acid + Choline

ATP

P
p70S6K

mTOR

CD

Deptor
mLST8

Raptor
PRAS40
P

Specic Inhibitors of mTOR Ac<va<on

Akt

EDL Muscle Undergoing Eccentric

Contrac'ons In Vitro
Akt

ONeil et al. J Physiol 587: 3691-3701, 2009

mTOR Ac<va<on by Muscle Contrac<on

Blocking mTOR ac<vity with rapamycin blocks
contrac<on-induced muscle protein synthesis (mTOR
ac<va<on required)
Blocking ac<va<on of PI-3 kinase/Akt with wortmannin
does not prevent ac<va<on of mTOR with muscle
contrac<on (IGF-1 ac<va<on not necessary)
Inhibi<on of TSC is not required for contrac<on induced
muscle protein synthesis (ERK 1/2 ac<va<on is not
necessary)
Blocking phospholipase D ac<vity does inhibit
contrac<on-induced ac<va<on of protein synthesis
(ac<va<on of phospholipase D necessary)

mTOR Ac<va<on by Muscle Contrac<on

Insulin
I R

IRS

PI3K

Ptdlns(4,5)P2

Ptdlns(3,4,5)P3

IGF-1

ERK 1/2
P
P TSC2

Phospha'dylcholine

Phospholipase D

TSC1

Akt
GDP

GTP
Rheb

Rheb

ADP

Phospa'dic Acid + Choline

ATP

P
p70S6K

mTOR

CD

Deptor
mLST8

Raptor
PRAS40
P

mTOR Ac<va<on During Exercise

I R

IRS

Insulin
PI3K

Ptdlns(4,5)P2

pO2
P

REDD1

TSC2

Ptdlns(3,4,5)P3

ATP/ADP: PC: Glycogen

TSC1

Akt

AMPK
GTP
Rheb

GDP
Rheb

AMPK: AMP ac<vated protein kinase

LKB1: Serine/threonine kinase 11
REDD1: Regulated in DNA damage and development CD

mTOR
Deptor

p70S6K

Blocking protein synthesis during exercise

conserves cellular energy to support muscle
contrac<on and work

LKB1

mLST8

Raptor
P

PRAS40

Does Feeding During Exercise Prevent Suppression of mTOR Ac<vity?

Rats were administered saline, glucose (135 mg/100g BW), or BCAA (135 mg/100g BW) by
oral gavage 30 min before exercise.

Murakami T et al. BBRC 405: 615-619, 2011

Sedentary

Exercised

Phosphoryla<on of 4E-BP1 and Expression of REDD1

Murakami T et al. BBRC 405: 615-619, 2011

Eect of Exercise on Muscle Protein

Synthesis

Muscle Protein Synthesis and Breakdown Before

and Amer Resistance Exercise
FSR and FBR (%/h)

Synthesis
Breakdown

0.15

b
c

0.10

a
B

0.05

A
0.00

24h
48h
Rest
3h
Means with dierent leders are signicantly dierent (p 0.05)
Dierences between protein synthesis and breakdown equals net protein balance

Phillips SM et al. Am J Physiol Endrino Metab 273:E99-E107, 1997

Protein or Amino Acid Supplementa<on

Following Resistance Exercise

L-Leucine

Experimental Design
Intravenous infusion of a balanced
amino acid mixture at rest and amer
a leg resistance exercise protocol
Exercise was knee extensions
Infusion started

Biolo et al. Am. J. Physiol. 1997

% Change from Basal!

400"

300"
200"

100"
0"

AA (Rest)"

AA (Post-exercise)"

Protein Synthesis!

Biolo et al. Am. J. Physiol. 1997

Rate of Protein Synthesis

Protein Synthesis and Degradation with

Increasing Protein with Meal

Synthesis

10

15

20

25

30

Protein Consumption (g)

35

Type of Protein for Post Exercise Supplement is Important

.25

FSR (% h-1)

.20

Rest
Exercise

*#
*#

.15

.10
.05
0

Whey

Tang et al. J Appl Physiol 107:987-992,2009

Soy

Casein

Eect of Exercise plus CHO on Muscle

Protein Synthesis

D-glucose

Muscle Protein Synthesis Following Exercise and

Protein/CHO

10 subjects performed both upper and lower body

resistance exercise las<ng ~ 1 hour
Subjects received a beverage volume of 2.5 ml/kg
every 30 min to ensure a given dose of 0.3 g/kg of a
casein protein hydrolysate per hour combined with
either:
0 g/kgh-1 carbohydrate (PRO treatment),
0.15 g/kgh-1 carbohydrate (PRO LCHO treatment),
or 0.6 g/kgh-1 carbohydrate (PRO HCHO
treatment).
Koopman R et al. Am J Physiol 293:E496-E504, 2007

0.14
0.12

FSR (%/H

0.10
0.08
0.06
0.04
0.02
0.00

PRO

Koopman R et al. Am J Physiol 293:E496-E504, 2007

PRO + LCHO PRO + HCHO

Studies Support the Use of CHO in a PRO

Supplement Post Exercise

Carbohydrate and Amino Acid Supplementation

Bird et al. EJAP, 97:225-238,

2006.
32 subjects trained for
12 weeks while
consuming several
different nutritional
interventions
Supplements consumed
during exercise and post
exercise:
Placebo
6% CHO solution
6 g EAA
CHO + EAA

Carbohydrate and Amino Acid Supplementation

Bird et al. EJAP, 97:225-238, 2006.

Carbohydrate and Amino Acid Supplementation

Bird et al. EJAP, 97:225-238, 2006.

Eects of exercise and insulin infusion on

protein synthesis and degrada<on
Protein
Synthesis
Without Insulin
Rest
Post Exercise
With Insulin
Rest
Post Exercise

307
6510
514
649

Protein
Degradation

*

*

Units are nmolmin-1100 ml leg volume.

Biolo et al. Diabetes (1999).

Net

468
7410

-16
-9

483
529

3
12

Carbohydrate and Amino Acid Supplementation

3-methyl-histine release

Bird et al. EJAP, 97:225-238, 2006.

mTOR Signaling Pathway

Exercise

Protein
Blood amino acids

Glycogen

p70S6K

PC

4EBP1
rpS6
FOXO3A

GSK-3

protein
degradation

ATP/ADP

AMPK

mTOR
Akt

PA

ribosomes

mRNA
translation
initiation

eIF2B

protein
synthesis

Net Protein Accre'on

Insulin

CHO

Protein Synthesis and Degradation with

Increasing Protein with Meal
Rate of Protein Synthesis
and Degradation

Synthesis
Degradation
Accretion

20
5

40
10

60
15

80
20

100
25

120
30

PlasmaConsumption
Insulin (U/ml)
Protein
(g)

35
140

Eect of Phospa<dic Acid Supplementa<on on

Resistance Exercise Training Adapta<ons
28 subjects were randomly assigned to one of two
treatment groups (14 subjects per group):
Placebo
Phospha<dic Acid (750 mg/d) from soy
Subjects completed an 8 week periodized whole-body
resistance training program
Tested for changes in body composi<on by DEXA, muscle
hypertrophy (CSA of the rectus femoris) and muscle
strength (1 RM for bench press and leg press)

Joy JM et al. Nutr Metab 11:29-39, 2014

Changes in CSA Rectus Femoris and Lean Body Mass

CSA
8

Lean Body Mass

75

PA
PLA

PA
PLA

Kg

cm2

60

Pre

Post

Pre

Joy JM et al. Nutr Metab 11:29-39, 2014

Post

45

Pre

Post

Pre

Post

Joy JM et al. Nutr Metab 11:29-39, 2014

Summary
mTOR is a serine-threonine kinase with two congura<ons
(mTOC1 and mTOC2). mTOC1 is sensi<ve to rapamycin and
responsible for cell growth and training adapta<on
mTOR is responsible for ac<va<on of transla<on ini<a<on and
therefore control of protein synthesis
Growth factors, nutri<on and muscle contrac<on can regulate
mTOR ac<vity
Growth factors ac<vate mTOR via the IRS-1/PI-3 kinase/Akt
pathway
Nutri<on (primarily L-leucine) ac<vates mTOR by ac<va<ng
Rag A/B, which recruits the mTOC1 complex to a lysosomal
membrane where it can interact with Rheb
Muscle contrac<on ac<vates mTOR by genera<ng
phospha<dic acid, which binds to the mTOR protein

Summary completed
During exercise, mTOR is inac<vated by ac<va<on of AMP
Kinase and increased expression of REDD1
Post exercise net protein balance is remains nega<ve un<l
nutrient interven<on
Providing protein and/or L-leucine will act addi<vely with
muscle contrac<on to increase muscle protein synthesis and
produce a posi<ve net protein balance
The addi<on of CHO to a post workout protein supplement
does not enhance muscle protein synthesis. However, it will
reduce the rate of protein breakdown and increase net
protein accre<on
Taking a phospha<dic acid supplement (750 mg) daily may
increase the rate of muscle and strength development during
a resistance exercise training program

Sports Nutri'on Workshops

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Topics include:
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