Lecture 5

Cell Biology, Structure and Extracellular Matrix

 The Hierarchy of Life
§ At each level of organization, novel properties emerge that were not present at the previous level

§ A group of cells and surrounding extracellular matrix that function together to perform one or
many specialized activities is called a tissue.
The Extracellular Microenvironment
 Functions of the Extracellular Microenvironment
§ Mechanical support for cell anchorage and tissue survival
§ Determination of cell orientation; control of cell growth, cell confinement
§ Maintenance of cell differentiation
§ Establishment of tissue microenvironment
§ Sequestration, storage, and presentation of soluble regulatory molecules
The ECM is secreted by the cells that reside within it

Fibroblasts surrounded by secreted collagen fibers

 Factors Regulating Cell Phenotype
§ Up or down-regulation of specific
pathways
- Integrins
- Growth factor receptors
- Modification of lipids

§ Production and secretion of matrix

components, GFs and hormones.
§ Never operate individually, always
tightly tied to internal and external
microenvironment of the cell
 ECM Composition and Function in Connective Tissues
(Bone, Muscle, Cartilage)
§ Fibrous proteins whose role is mainly structural:
– Collagen gives tissues tensile strength
– Elastin gives tissues elasticity/resilience

§ Fibrous proteins whose role is mainly adhesive:

– Fibronectin: promotes the attachment of fibroblasts and other cells to the ECM
– Laminin: promotes the attachment of epithelial cells to the basal lamina

§ Glycosaminoglycans (GAGs) and proteoglycan molecules:

– Form a highly-hydrated gel-like substance in which fibrous proteins are embedded
– The polysaccharide gel resists compression
– The aqueous phase of the gel permits the rapid diffusion of nutrients, metabolites, and hormones

§ Growth factors and cytokines

Collagen: the fibrous proteins of the matrix
Collagen
§ Most abundant protein present in the
human body (25%)
§ Major component of skin and other
musculoskeletal tissues
§ Collagen is a rod-type polymer; 300 nm
long with a Mw of 300kDa.

§ At least 28 different types identified in humans

§ Type I – IV, 80-90%
§ Structure preserved across species; little immune response when transplanted
§ Type I collagen
§ Single most abundant and most thoroughly studied
§ Three polypeptide chains with similar AA compositions arranged in a triple helix
Periodic Structure of Collagen : Gly–X–Y repeats
§ X is typically proline.
§ Y can be any amino acid,
but is often hydroxyproline.
§ The small size of glycine
(side chain is just a
hydrogen atom) enables
the 3 helices to pack tightly
together.
§ Each α-chain contains
about 1000 amino acids
and is about 300 nm long.
 Collagen Types and Structure
§ Collagen I: skin, tendon, vascular ligature, § Primary: ⅓ of residues are glycine;

organs, bone (main component of the organic ¼ of residues are proline or

part of bone) hydroxyproline

§ Collagen II: cartilage (main component of § Secondary: triplet sequence (Gly-

cartilage) Pro-Hyp) for helix stabilization

§ Collagen III: reticulate (main component of § Tertiary: triple helix 300 nm long
reticular fibers), commonly found alongside
§ Quaternary: formation of
type I.
microfibrils; quasi-hexagonal lattice
§ Collagen IV: forms bases of cell basement
membrane

§ Collagen V: cell surfaces, hair and placenta

 Collagen fibers

AFM images of the collagen fibers

Collagen fiber in SEM
 Collagen gels
• Dissolve collagen in acid solution and then change the pH or heat to form a gel --
very easy to do and amenable to many process and applications.
• Basic studies
• Commercial devices -- hemostatic devices - stop bleeding
• Sutures, blood vessels, heart valves, burn dressings, nerve regeneration, meniscus
repair, etc.
 Collagen vs. Gelatin

Also enzymatically degradable by collagenases and metalloproteinases à Control over degradation rates
 Collagen
Collagen-like peptide (CLP)
§ Main component of extra cellular matrix (25-35% of
human body proteins) § Short synthetic peptide
§ Mimic triple helix
§ Widely found in tendon, skin, cornea and cartilage § Gly-Xaa-Yaa tripeptide motif
§ 29 different types
§ Mediating cell adhesion, migration, tissue § Study triple helix stability
scaffolding and repairing § Mimic collagen fibril formation

1. Kadler, K. E.; Baldock, C.; Bella, J.; Boot-Handford, R. P.. J. Cell Sci. 2007, 120, 1955.
2. http://www.proto-col.com/blog/2014/07/collagen/ 15 J. 2013, 49, 2998.
3. Luo, T. Z.; Kiick, K. L. Eur. Polym.
 Triple helix hybridization

Above Tm

Yu, S. M.; Li, Y.; Kim, D. Soft Matter 2011, 7, 7927. Urello, M. A.; Kiick, K. L.; Sullivan, M. O., J. Mat. Chem. B
2014, 2, 8174-8185.

§ CLP as building block to develop

amphiphiles
§ Self-assembly into nanoparticles
§ Targeted drug delivery

Li, Y.; Foss, C. A.; et al., P Natl Acad Sci 2012, 109, 14767.
 CLP assembly
Lateral electrostatic
π-π stacking Metal ligand coordination
interaction

Cejas, M. A.; Kinney, W. A.; et al., J Am Pires, M. M.; Chmielewski, J., J Am Chem Soc 2009, 131, 2706.
Rele, S.; Song, Y. H.; et al., J Am Chem
Chem Soc 2007, 129, 2202. Pires, M. M.; Ernenwein, D.; et al., Biomacromolecules 2011, 12,
Soc 2007, 129, 14780-14787.
Cejas, M. A.; Kinnney, W. A.;et al., P Natl 2429-2433.
17 Acad Sci USA 2008, 105, 8513-8518. Pires, M. M.; Lee, J.; et al., Langmuir 2012, 28, 1993-1997
 CLP conjugate assembly
C15
A5K4G(GPO)3GFOGER(GPO)3G

Krishna, O. D.; Wiss, K. T.; Luo, T. Z.; Pochan, D. J.; Theato, P.;
Luo, J. N.; Tong, Y. W. Acs Nano 2011, 5, 7739.
Kiick, K. L. Soft Matter 2012, 8, 3832
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Organelles: Peroxisomes
• Break down fatty acids, branched chain fatty acids, D-amino acids,
polyamines
• Synthesize plasmalogens
• Plasmalogens - phospholipids critical for the normal function of
mammalian brains and lung

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