Group A Sara Smith & Anna Mitchell October 15, 2012

The path of dietary fats

The function of adipocyte LPL (aLPL)
The effects of aging, exercise, & sex on aLPL
The influence of aLPL on body composition & fat

distribution
The handling of different types of dietary fats
The formation of lipid droplets

The white adipose tissue

 Typical American meal: 100 g carbohydrate,

30-40 g fat The amount of TG in circulation is ~3 g. Does a meal raise the plasma TG 10-fold? NO! The plasma TG will rise less than 100%.
So.how does this process work?

Frayn, 2002

What Path Does a Quadruple Cheeseburger Take on its Way to Becoming a Lipid Droplet?

Houston, 2006, pg. 149

After a fatty meal, the triglycerides (TG) are formed from the fatty acids in the intestinal cells.

The TG combine with protein and phospholipid to form a chylomicron. Chylomicrons are one type of lipoprotein.
Houston, 2006

 Function- transport dietary fat and cholesterol

through circulation. CHYLOMICRONS: Produced in the intestinal cell

Largest of all lipoproteins

90% TG, 8% PL, 5% Cholesterol

Apo AI, AII, B-48, CII, CIII, E

Gropper & Smith, 2012

Adipose tissue picks up the fatty acids (FA) liberated from chylomicron by lipoprotein lipase (LPL) FA transport into the cell is accomplished using CD36 The FA diffuse into interstitial space, no specificity LPL is bound on surface of capillary endothelial cells Chylomicron binds to receptor Held in place while LPL works Houston, 2006; Julve et al., 2008 Apo CII, CIII regulate LPL activity

Albumin

Chylomicron Fatty acids

Insulin

CD36

IR

CD36
PI3K

+
nucleus

Control of CD36 activity

 Ligands for receptor binding. Receptor mediated

binding removes lipoproteins from circulation. Activators of enzymes: transfer lipids to and from lipoproteins.
Apo B-48: Plays an essential role in the intestinal absorption of dietary fats Apo C-II: activates lipoprotein lipase (LPL) Apo C-III: inhibits lipoprotein lipase (LPL); regulates activity of Apo C-II.

Julve et al., 2008; Gropper & Smith, 2012

 Overexpression of Apo A-

II in mice increased postprandial plasma TG concentrations Presence of Apo A-II in HDL might influence HDL apolipoprotein composition or apolipoprotein transfer to triglyceride-rich lipoproteins (TRL), thereafter altering TRL postprandial catabolism.
Increased content of Apo

A-II was associated with decreased content of Apo C-II and Apo C-III.

 Depleted chylomicron = chylomicron remnant Chylomicron remnant taken up by liver Receptor mediated endocytosis The life span of each chylomicron is 10 min.

Julve & Smith, 2012

 Intracellular lipolysis in adipose tissue is suppressed by

insulin No need for hydrolysis of TG into FFA when there is adequate energy (post-prandial state)
Negative valuefat mobilization Postive valuefat deposition

Bickerton et al., 2007; Frayn, 2002

1.

2.
1.

Insulin suppression of nonesterified fatty acids (NEFA) release from adipose tissue Clearance of TG in adipose tissue increases in post-prandial period
Insulin upregulates adipose tissue expression of LPL.

3.

In post-prandial state, HSL is suppressed by insulin and the pathway of FA esterification is stimulated, and LPL-derived FA are drawn into the adipocyte by the resultant concentration gradient
Frayn, 2002

 Synthesis:
Synthesized in adjacent fat

cells Secreted from the cell Attached to the endothelial lining of a nearby capillary
Role:
Free FA from lipoproteins so

that the FA can flow down a concentration gradient into adjacent adipocyte Requires cofactor, Apo C-II, to be fully active
Houston, 2006; Wang and Eckel, 2009

 Results suggest that

chylomicrontriglyceride is the preferred substrate for LPL. There is a preferential channeling of FA derived from LPLmediated chylomicron into adipose tissues

 Plays a key role in bringing

TG from chylomicrons into the adipose tissue Binds LPL and chylomicrons Responsible for the transport of LPL into the capillaries and actively transports LPL across endothelial cells

Davies et al., 2008

 An ER membrane protein involved in the

posttranslational folding and/or assembly of LPL into an active enzyme

Peterfy, 2011

 1.Depot differences in LPL activity, which is a rate-

determining step in the uptake of circulating TG 2.Females preferential post absorptive direct FFA uptake by subcutaneous adipose tissue 3. Males enhanced postprandial meal-derived FFA uptake by visceral adipose tissue.

Differences in Fat Distribution

Male

Female

 Women have more body fat than men, but in contrast to the deleterious metabolic consequences of the central obesity typical of men, the pear-shaped body fat distribution of many women is associated with lower cardiometabolic risk. ( Karastergiou) body fat is stored in the adipose organ which consists of definable fat depots Subcutaneous white adipose tissue (SAT) depots, just under the skin, store ~80-90% of total body fat, mainly in the abdominal (around the waist), subscapular (on the upper back), gluteal and femoral (thigh) areas. (Karastergiou)

Fat distribution is modulated by sex steroids . Gluteofemoral adipocytes of women are more efficient in storing FA via the direct pathway and also show higher LPL activity.

BAT

 Menopause is followed by a reduction of adipose tissue

towards a more android phenotype. Importantly, its visceral adiposity that rises in women during the premenopausal transition, presumably due to the fall in estrogen levels (Karastergiou,2012)

 LPL activity shows a strong positive relationship with

regularly performed exercise (Houston, 2006).

 LDs are storage organelles, but they also

have a role in maintaining homeostasis and metabolism

LD Composition:
Organic core of TG and sterol esters bounded

by a layer of phospholipids Several types of proteins Structural proteins (perilipin family) Lipid-synthesis enzymes (ex. Acetyl CoA carboxylase) Lipases (ATGL) Membrane-trafficking proteins (Rab5)

Wang and Sztalryd, 2011; Guo et al., 2009

Lipid Droplet Formation

FA are carried extracellularly by albumin and lipoproteins enter cells. FA are released from TG in lipoproteins by LPL and enter cells by passive diffusion facilitated by FA transport proteins (CD36)
Guo et al., 2009

FA enter a bioactive pool through conjugation to CoA, forming fatty acyl-CoA, in an energy requiring reaction

Guo et al., 2009

FA acyl CoA diacylglycerols TAG or Phospholipid Takes place in the ER

Guo et al., 2009

Lipids accumulate and form LDs. How does this occur?

Guo et al., 2009

I. Neutral lipids form a lens of oil in the ER bilayer that subsequently buds from the membrane, taking with it the phospholipids from the cytosolic leaflet. Guo et al., 2009

II. Neutral lipids accumulate between the leaflets of the ER membrane but, instead of budding, emerging LDs are excised from the membrane, taking with them phospholipids from both the cytosolic and luminal leaf. Guo et al., 2009

III. Small bilayer vesicles that remain tethered to the ER membrane are used as a platform for making LDs. Newly synthesized neutral lipids are pumped into the vesicle bilayer and fill the intermembrane space, eventually squeezing the vesicular lumen so that it becomes a small inclusion inside the LDs. Guo et al., 2009

 Perilipin: binds to LD

surface. Acts as a scaffolding protein for key lipolytic proteins. May retard TAG hydrolysis through sterically blocking access of HSL to the droplet surface. Also regulates lipolysis by phosphorylation/ dephosphorylation events that induce changes in protein interactions at the LD surface.

Wang, 2011; Brown, 2011

 Phosphorylation of perilipin simultanesouly triggers:

Release of CGI-58, a co-lipase essential for Adipose

Triglyceride Lipase (ATGL) activation ATGL gains access to TAG Dispersion of CGI-58 reveals docking site for HSL

Wang, 2011; Brasaemle, 2010

 Function: To store excess energy

WAT composes as much as 20% of the body weight in

men and 25% in women WAT cells contain a single large fat droplet, which forces the nucleus to be squeezed into a thin rim at the periphery.

 Have receptors for insulin, growth hormones,

norepinephrine, and glucocorticoids

Upon release of insulin from the pancreas, WAT cells

insulin receptors cause a dephosphorylation cascade that lead to the inactivation of HSL. Upon release of glucagon from the pancreas, glucagon receptors cause a phosphorylation cascade that activates HSL, causing the breakdown of stored fat to FA, which are exported into the blood and bound to albumin, and glycerol, which is exported into the blood freely.

 Dietary fat is transported in chylomicrons

LPL, with the help of Apo C-II, releases fatty acids

from chylomicrons
LPL expression varies between sexes LPL expression increases with exercise LPL expression changes with age

Fatty acids use membrane transporter CD36 to enter

adipocytes Within the adipocytes, fatty acids form lipid droplets Differences in dietary fat types influences how the fat is handled. WAT is primarily a fat storage cell

 DL Brasaemle (2010). Lipolysis Control: the plot thickens. Cell Metab,

11. 173-174. DA Brown (2011). Lipid droplets: Proteins floating on a pool of fat. Current Biology, 11. R446-R449. BSJ Davies, H Waki, AP Beigneux, E Farber, M Weinstein, DC Wilpitz, L Tai, RM Evans, LG Fong, P Tontonoz, & SG Young. (2008). The Expression of GPIHBP1, an Endothelial Cell Binding Site for LPL and Chylomicrons, is induced by PPARy. Mol Endocrin, 22. 2496-2504. Gropper S.S.and J.L. Smith. Advanced Nutrition and Human Metabolism.(6th edition 2012): Wadsworth, Belmont, CA (ISBN 13: 978-1-133-10405-6) M Peterfy (2011). Lipase maturation factor 1: a lipase chaperon involved in lipid metabolism. BBA, 1821. 790-794. H. Wang and R.H. Eckel. (2009) Lipoprotein lipase: from gene to obesity. Am J Physiol-End, 297. E271-E288.