Introduction

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In order to form complex structures (e.g. tissues, organs, etc) cells must
interact with one another
This requires Cell Adhesion Molecules, which are typically proteins
Occurs in the Cell Junction
Now let's review a schematic representation of the Adhesive Proteins. 

Types of Cell-Cell Junction

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Many transmembrane proteins are involved in cell-cell interactions:

Cadherins
Mucin-like CAMs
Integrins
Now let’s review a schematic of the Cell-Cell Junction. 
 Cadherins I
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Cadherins (named for “Calcium-Dependent Adhesion") are a class of type-

1 transmembrane
proteins.
They play important roles in cell adhesion, forming adherens junctions to
bind cells within tissues
together.
The cadherin family consists of three different proteins, all of which
prefer homophilic interactions
– that is, they bind to the same type of protein (e.g. E-cadherin binds E-
cadherin)
Cadherins are Type I membrane proteins
Cadherins are Calcium-dependent; require Ca 2+ to function
E-cadherin: 
the most abundant cadherin
Found in the pre-implantation embryo as well as in non-neuronal
epithelia
Holds epithelia together
Now let's review a schematic of E-Cadherin in both insects and
vertebrate. We will learn more about membrane proteins later this
week. 

Cadherins II
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P-cadherin:  found in the trophoblast, required for nidation
N-cadherin:  found in neuronal tissue, lens, heart, and skeletal muscle
N-cadherin, also known as Cadherin-2 (CDH2) or neural cadherin (NCAD)
is a protein that in
humans is encoded by the CDH2 gene
Changes in cadherin expression often accompany metastasis!
Recall that metastasis is the process whereby cancerous cells break free
from the primary
tumor and spread
Reduced cell adhesion is essential for metastasis
Now let’s review a schematic of N-Cadherin as shown below. 

Important Note:
Nidation is the process by which an embryo burrows into the endometrium of the
uterus and is
also called implantation. 
NCAMs
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N-CAMs, also known as Nerve-Cell Adhesion Molecules or CD5 is
a homophilic binding glycoprotein expressed on the surface of
neurons, glia, skeletal muscle and natural killer cells.
Encoded by a single gene
a glycoprotein of Immunoglobulin (Ig) superfamily
Alternative splicing generates 3 isoforms
NCAM 180 (long cytoplasmic domain)
NCAM 140  (short cytoplasmic domain)
NCAM120 (GPI anchored) has lost the transmembrane domain and is
instead anchored to the membrane by glycosyl phosphatidylinositol
Function in learning and memory!                                                                      
Now let’s review a schematic of N-CAM1 as shown below.      
 Mucin-like CAMs
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Mucin-like CAMs are hetrophilic

Mucins are a group of serine and threonine rich proteins and they are
heavily glycosylated
Two mucin-like molecules (CD34 and GlyCAM-1) on certain endothelial
cells of lymph nodes bind to L- selectin onleukocytes
 PSGL-1 is a mucin-like molecule on neutrophils. It interacts with E-
selectin and P-selectin on inflammed vascularendothelium
Prefer binding to different proteins:  selectins
Selectins have a lectin binding domain
Important in leucocyte/tissue interactions, helping them pass through
blood vessel walls
Rich in carbohydrates
Now let’s review a schematic of Mucin-like CAMs as shown below. 

Integrins
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Integrins are transmembrane receptors that facilitate cell-extracellular

matrix (ECM) adhesion:
Are also heterophilic
Heterodimers with an alpha and beta subunit
Primarily bind extracellular matrix (ECM) proteins such as fibronectin or
laminin
Integrin expression is stimulated by Platelet Activating Factor (PAF),
which is excreted by blood vessel endothelialcells
Now let’s review the structure of Integrins as shown below.
 Types of Cell Junctions
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The various CAMs we have covered are all involved in different types of
cell to cell junctions:
Tight Junctions
Gap Junctions
Cell-Cell Junctions
 Tight Junctions
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Tight junctions connect epithelial cells and prevent the movement of

fluids from one side of the epithelium to the other
Fluids and other essential molecules must therefore pass through the cells
osmotically or actively if they are to move from one side of the
epithelium to the other
E.g. stomach and intestinal lining
Note the components of the protein complex linking together the cells
Now let’s review a schematic of Tight Junctions as shown below. 
                            
Gap Junctions
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Gap Junctions are distributed on the lateral surfaces of cells and allow the
exchange of small
molecules between neighboring cells
They are actual connections/portals between the cells but are highly
regulated
The cytosols of the neighboring cells are linked by the hydrophilic
channels, which are themselves
comprised of transmembrane proteins called Connexin, which aggregates
into Connexons.

Now let's review a schematic of Gap Junctions as show below.

Gap Junctions (cont'd)
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These are plasma membrane specializations that contain channels which

allow for the
transportation of low molecular weight substances between the cytoplasm
of neighboring cells
Each gap junction channel contains a hemi-channel that is situated in
appositional membranes
Each hemi-channel consists of six subunit proteins called connexins
Sequencing of the human genome showed humans have 14
distinct connexins, subsets of
which are expressed in all of the tissues of the body
 Cell-Cell Junctions
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Cell-cell junctions tend to have structural roles

They hold cells together by forming a “bridge” between their individual
cytoskeletons
Two different types:
Adherens junctions
Desmosomes
Adherens Junction
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Defined as a cell junction whose cytoplasmic face is linked to the
actin cytoskeleton
May form bands encircling the cell
Alternately may form as spots of attachment to the extracellular matrix
Important Note:
Notice the connection of Cadherin E to the actin cytoskeleton) via alpha and
beta catenins.
Now let's review a schematic of the Adherens Junction as shown below. 

Desmosomes
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Desmosomes are formed
when desmocollin and desmoglein (both cadherins) are cross-linked to
keratin filaments, forming a cytoplasmic plaque
The keratin (a form of IF, recall) helps distribute forces between layers of a
tissue due to their full
permeation through cells
Now let's review a schematic of Demosomes as shown below. 

Cell motor proteins

Introduction
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 Some of the more interesting systems of proteins in the human body
are those which help form cellular  motors. Cellularmotors are involved
in:
Muscular contractions (both smooth and skeletal)
Intracellular transport
Extracellular transport
Movement of chromosomes during mitosis/meiosis
Cellular motility
Now let’s review a schematic of the Cell Motor Proteins as shown
below. 
 Myosin is a superfamily of genes
13 different genes in humans but only 3 are well characterized:
Myosin I
Myosin II
Myosin V
All myosin isoforms have an N-terminal head and a C-terminal tail
The myosin “tail” is a coiled coil of two alpha helix
The head is comprised of two identical units containing a heavy chain
and two light chains
The light chains will bind Ca2+
This structure is critical to the motor function of myosin!
Now let’s review a schematic of the General Myosin Structure as
shown below. 

Myosin I & II
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The Myosin I protein serves a very different purpose from the Myosin II

we’ve been looking at so far.
Cellular structure & organelle motility
Stabilizes microvilli by linking the plasma membrane to the actin bundles
Co-purifies with Golgi vesicles, which suggests that it is strongly bound
to golgi and is involved in
their movement along actin microfilaments
The Myosin II protein is involved in muscular contraction. There are
three classes of muscle:
Skeletal
Smooth
Heart
Now let’s review a schematic of the Myosin at (a) Relaxed
state  & (b) Contracted state. 
 Myosin I, II & V Structures
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While the myosins are in general very similar, their differential

functions derive from the differences in their structures
Myosin II and V are very similar, with differences primarily the light chain
region
Myosin 1 has a single alpha helical coil tail, which is significantly shorter,
and only a single head (motor domain)
Myosin V appears to be involved in the transport of exocytotic vesicles to
the plasma membrane
These vesicles typically contain materials to be exported from  the cell
Thus Myosin V is involved in extracellular transport
 Skeleton Muscle
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Able to contract rapidly but tires quickly

Comprised of an organized complex of actin and myosin (actomyosin)
Now let's review a schematic of the Skeleton Muscle as shown below. 

Important Note:
The actomyosin complex is visibly striated under a microscope.
Skeleton Muscle Action I
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Now let’s review the Actin-Myosin interactions as depicted in the

schematic below.

 
Important Note
Notice that the action of actin and myosin acts in a ratchet-like fashion,
where one ATP is consumed for each individual ratcheting of the
myosin head against the actin filament per actomyosin interaction.
Skeleton Muscle Action II
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Troponin and tropomyosin wind around the actin filaments

The organized structure of these individual filaments comprises
the actual skeletal muscle fibers
 Smooth Muscle
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Smooth muscle is involuntary and non-striated:

Does not form ordered arrays like skeletal muscle
Lines the walls of hollow organs
E.g. urinary tract, digestive system, reproductive organs, etc..
Contractions are smoother and slower
Regulated by autonomic nervous system as well as hormones and
growth factors
Smooth muscle contains the protein caldesmon instead of troponin
Ca2+ dependent phosphorylation of caldesmon and myosin light chains
causes contraction
Now let's review a schematic of the smooth muscle depicted below.

Heart (Cardiac) Muscle

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Cardiac muscle is involuntary like smooth muscle, but striated like

skeletal muscle.
Powerful and strong
Like skeletal muscle, composed of actin & myosin
actin filaments are thin, causing lighter appearance to the actin bands
myosin filament is thicker, lending a darker appearance to the  myosin
bands
 cardiac muscle cells are typically branch-like instead of linear.
Basic contractile function is similar to skeletal muscle
Now let's review a schematic of the Cardiac muscle as depicted below. 
Intracellular Transport
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 Living cells have been observed to have organelles and vesicles moving
long distances in straight lines
This is non-random motion or directed transport
E.g. transport of proteins from ribosome to synapse in neural cells
This transport has been shown to be protein-dependent and occurs in
both anterograde
(center of the cell towards the periphery) and retrograde (periphery of the
cell towards the
center) directions
Anterograde Cellular Transport
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Anterograde cellular motion along microtubules is kinesin-dependent

Kinesin is a protein with 14 different isoforms in humans
Similar to myosin
Now let’s review a schematic of the protein Kinesin I. 

Kinesin
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380kDa protein
2 heavy chains
Heavy chain n-terminal head has an ATPase-domain and a tubulin
binding domain
C terminal ends wrap around each other in a coil
2 light chains
Binds microtubules and “ratchets”, consuming ATP
Now let's review a schematic of the Kinesin Chart. 

Retrograde Cellular Transport

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Retrograde cellular motion along microtubules is dynein-dependent

Dynein is composed of four different subunits
Total mass of 1.5MDa
2 heavy chains with ATPase activity
2 intermediate chains
Several light chains
Cargo binding domain
Dynein
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 Dynein is a family of cytoskeletal motor proteins that move along
microtubules in cells
Convert the chemical energy stored in ATP to mechanical work 
Transports various cellular cargos, provides forces and displacements
important in mitosis
Drives the beat of eukaryotic cilia and flagella
Retrograde cellular motion along microtubules is ATP and dynein-
dependent
Dynein is composed of four different subunits
 Total mass of 1.5MDa
2 heavy chains with ATPase activity
2 intermediate chains
Several light chains
Cargo binding domain
Now let’s review a schematic of the Dynein cellular motion along
microtubules.
 Cellular Motility
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Protozoans are not the only cells that are motile

Sperm cells are propelled by cilia and flagella
Epithelial cells in the lungs use cilia to remove contaminants
Nearly every vertebrate cell has a cilium usually they are nonmotile and
carry receptors for
signaling
Retinal rod/cone cells have an outer segment connected to the cell body
by cilium
Olfactory neurons use cilia to carry G-protein chemoreceptors
Cilia and Flagella
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Cilia and flagella are similar in structure

Cilia are shorter and occur in larger numbers than flagella
A eukaryotic flagellum is a bundle of nine fused pairs of microtubule
doublets surrounding two central single microtubules
The so-called "9 + 2" structure is characteristic of the core of the
eukaryotic flagellum called an Axoneme
9 tubulin doublets surrounding 2 central tubulin singlets
Doublets are linked by nexin bridges, contain dynein arms
Now let’s review the cross section of an Axoneme. 

Mitotic Spindle Apparatus

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Mitotic Spindle Apparatus is responsible for the separation of
chromosomes during cellular division
The MSA is composed of at least 795 proteins
While a full review of mitosis/meiosis is beyond the scope of this course, it
is important to understand
that the underlying structural and motor proteins are the same as those we
have already
examined. e.g. microtubules, kinesin, dynein, etc..
Now let's review a schematic of the Mitotic Spindle Apparatus as shown
below.

Transmembrane proteins

Introduction
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Membrane proteins are important in terms of signaling and transport:

Some of the most important transport functions involve ion channels,
such as those used in the electronic transport chain, which is essential
for metabolism and regeneration of ATP
 Integral membrane proteins are permanently attached to the cell
membrane:
Transmembrane, or polytopic, proteins span the entire width of the
membrane at least once, from the cytosol to the cell surface
Monotopic proteins are attached only to one side of the cell
membrane
Transmembrane Proteins
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Typically have one of two architectures:

Helix bundle (found in all membranes)
Primarily alpha helix structures
Beta Barrel (found only in bacterial or mitochondrial membranes)
Primarily beta-sheet structures

Transmembrane Proteins
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Not just transport across the primary cell membrane!

Also includes transport across mitochondrial membrane or endoplasmic
reticulum
Unique structure is required for this function:
Amino acids in the region that spans the membrane must have exposed
hydrophobic R groups, matching the environment of the membrane
Amino acids in the regions outside of the membrane must have exposed
hydrophilic R groups
Now let's review a schematic of Transmembrane Proteins as shown below. 

Hydropathy Plots
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 In Week 4 we will look in depth at some of the ways we can analyze
proteins in the lab
One way is a hydropathy plot
Looks at the hydropathic value of the R group for each amino acid, in
order, and plots it
Can often be used to predict structure, function, or location
Transmembrane proteins have a unique hydropathy plot profile
Now let's review a schematic representation of the Hydropathy plot. 

Monotopic Proteins
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Can be attached to the membrane in four different ways:

 Peripheral Membrane Proteins
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Peripheral membrane proteins are only temporarily attached to the

membrane, or to integral membrane proteins
This can include monotopic proteins that do not stay permanently
attached
Can also include proteins that bind integral membrane proteins
Translocon
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The translocon (commonly known as a translocator or translocation
channel) is a complex of
proteins associated with the translocation of polypeptides across
membranes
Any protein that is destined to be bound to a membrane typically has one
or more signal peptide
regions
The transport process begins during translation of the protein, causing
the
robosome/RNA/nascent protein complex to be bound to what is called
the Translocon
The translocon is a complex of proteins that assists in the transport of a
nascent protein (one
being synthesized) to, or across, the membrane
There is always a target AA sequence that helps direct the nascent
polypeptide to the appropriate
location
We can classify membrane proteins based on their target sequence
Membrane Proteins
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Now let’s review the various Membrane Protein types.

 Glycosylation
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The majority of secreted and transmembrane proteins are bound to

sugars – this is called glycosylation
Can aid in protein folding
Can act as a targeting signal to direct proteins to a specific intracellular
compartment
Can serve as receptors for cell-cell signaling
Can serve as binding sites for pathogens
Now let's review a schematic representation related to the different types
of Glycosylation. 
 N-Glycolysation & O-Glycolysation
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Sugars can be bound to proteins in two ways:

O linked:  bound to the OH groups of SER, THR
N Linked:  bound to the amino group of ASN
A protein can have  both O and N linked sugars
Protein/sugar complexes are also referred to as Oligosaccharides
N-linked oligosaccharides are bound to proteins as a preformed block (rather than one
sugar at a time, as with O-linked)
This process can involve many steps, including modification and trimming of the
sugar residues to
result in a final glycoprotein
O-linked oligosaccharides are produced in the Golgi apparatus by adding
one sugar at a time to the
protein, and results in a “sugar tree” complex where more than one sugar
may be added to a specific
site 
The reaction is mediated by membrane-bound glycosyltransferases,
which are specific for both the
acceptor AA and the donor sugar