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Protein surface interactions1

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4 views

Protein surface interactions1

Uploaded by

anitangabire017
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPTX, PDF, TXT or read online on Scribd
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PROTEIN - SURFACE

INTERACTIONS

BY: JJUUKO GEORGE


WILLIAM
PROTEIN – SURFACE INTERACTIONS

• Proteins interact with other molecules or entities on their outer


surfaces in various ways
• Interactions are crucial for many biological processes such as
• Protein-protein, protein-ligand, protein-membrane, protein-
carbohydrates interactions
• Protein surface interactions are responsible for biocompatibility of
medical devices or the lack thereof
• Important for designing devices that are biocompatible
• Biocompatibility is influenced by determining whether proteins
adsorb to the material’s surface
ADSORPTION VS ABSORPTION

Absorption is the process in which a


fluid is dissolved by a liquid or a solid
(absorbent).

Adsorption is the process in which


atoms, ions or molecules from a
substance (it could be gas, liquid or
dissolved solid) accumulate (adhere)
to a surface of the adsorbent (without
penetrating the surface).

Adsorption is a surface-based process


where a film of adsorbate is created
on the surface while absorption 3
PROTEIN–SURFACE INTERACTIONS

• Biomaterials’ surfaces are not bioactive


themselves
• Surface bioactivity is provided by the proteins
that adsorb to the surface
• Biomaterials form a chemical bond with the
tissues
• Involves the formation of a layer of
hydroxyapatite (HA) on the surface
• HA is a mineral component of natural bone,
and its presence on the surface promotes
surface integration with the surrounding bone
tissue
PROTEIN – SURFACE
INTERACTIONS
• Proteins rapidly
adsorb onto the
surface of the
biomaterial when it
comes in contact with
a body fluid
containing soluble
proteins
• Proteins saturate the
surface
PROTEIN – SURFACE INTERACTIONS
• Living cells do not actually contact the molecular structure of the material
surface itself
• Contact and interact with the molecular structure of the adsorbed protein layer
• Cells do not see the adsorbed protein layer
• Interrogate surroundings by way of membrane-bound receptors
• Bind to specific bioactive features presented by the adsorbed proteins
• Receptor–protein binding events are transduced through the cell membrane
• Stimulate specific intracellular processes that determine a cell’s response
• Cellular response is controlled by controlling the type of bioactive sites
• Bioactive sites are determined by
• Amounts and types of proteins adsorbed
• Types and surface density of the bioactive sites that may be available for cell interactions
• Orientation, conformation, and packing arrangement on the biomaterial surface
• Bioactive sites are presented in a manner such that they can be recognized by the
membrane-bound receptors
PROTEIN
STRUCTURE
• Complex copolymers made up of
four levels of structure
• Primary, secondary, tertiary, and
quaternary
• Primary structure involves the
specific sequence of the 20 L-amino
acids coded for by the DNA of a cell
• A given amino acid is referred
to as a peptide residue with a
general building block –NH–
CaHR–CO–
• R designates a specific side-
group structure that gives the
residue its specific functional
characteristics.
• Polypeptide chain formed by the
primary sequence is organized into
three basic types of secondary
structure
• α-helices, β-sheets, and loops
that connect helix and sheet
elements
PROTEIN STRUCTURE

• Secondary elements are organized together to form the tertiary


structure
• More than one polypeptide chain can be organized together to
form quaternary structure
• Each individual chain has a separate beginning (N-terminus) and
ending (C-terminus)
• Typically positively and negatively charged, respectively
• The specific functional group character of the peptide residues
combined with their spatial organization creates specific
bioactive domains in a protein’s structure that enable it to
perform its specific biological function.
PROTEIN STRUCTURE

• A protein is able to fold into its native state and maintain in that
state in aqueous solution
• Reduction in free energy due to decrease in the solvent
accessible surface area of the nonpolar residues in the
protein’s structure
• Hydrophobic effects structured with their hydrophobic residues
buried within the core of the protein and their hydrophilic
residues (charged and polar) lining the protein’s solvent
accessible surface
• You can have hydrophobic residues on the surface and
hydrophilic residues buried within the protein’s core
• Protein’s surface is highly amphiphilic
• Each charged residue on the protein’s surface has a designated
Protein Adsorption Behavior

• Single-Component Protein Solutions


• Multicomponent Protein Solutions
Single-Component Protein Solutions
Protein molecule in solution (P)
adsorbs to the material surface P . S
in a reversible manner with forward
and reverse reaction rate constants,
kf and kr
Orientational or conformational
changes transition adsorbed protein
to an irreversibly adsorbed state
Further reduces the free energy of
the system, described with a
reaction rate constant ki
Release of structured water from the
protein and surface back to bulk
water represents an important
factor that greatly influences these
processes
Single-Component Protein Solutions

• Described by the following chemical reaction equation:

• Where;
• P, S, P . S – protein concentration in solution, concentration of available
surface area sites for adsorption, and concentration of reversibly
adsorbed protein on the surface
• Water is complexed to the protein and surface, and then released to
bulk solution during the various stages of adsorption
• With sufficient interaction time, surface is inevitably saturated with
adsorbed protein
• State final protein layer is determined by manner in which proteins
organize themselves on surface
Single-Component Protein Solutions

• Factors that influence protein organization on surface


• Protein–surface interactions
• Protein–protein interactions
• Proteins that adsorb to the surface in latter stages of this process
are inhibited from the irreversible step by steric restrictions by
previously adsorbed proteins
• The final fraction of the adsorbed protein layer is more likely to be
maintained in its reversibly adsorbed state
• This can also be caused by a nonhomogeneous surface
Surface exposed to
single-component
protein solution and
rinsed by a buffer
• A - Proteins adsorb to random
areas of the surface in a
reversible fashion
• B - Adsorbed proteins reorient
and spread out to become
irreversibly adsorbed while new
protein molecules continue to
adsorb to open surface areas
• C - Process continues until most
of the available surface area is
covered with irreversibly
adsorbed proteins
• Last proteins to adsorb are
sterically prevented from
reorienting or spreading on the
surface by the previously
adsorbed proteins
• D – Exposure of surface to pure
buffer solution desorbs the
Multicomponent Protein Solutions

• Protein solutions with more than one protein


• Competitive process between the different proteins for adsorption to
the surface
• Major factor that influences adsorption of proteins is mass transfer rate
• Mass transfer rate of a given solute molecule to a surface is directly
related to its solution concentration and inversely related to its
molecular weight
• More concentrated and smaller proteins adsorb to the surface first
• Displaced by larger, more strongly interacting proteins
• An irreversibly adsorbed protein doesn’t desorb under flow of pure
buffer solution
• It is displaced from the surface by another more strongly interacting
protein that can more successfully compete for the same surface
functional groups
Vroman effect
• Vroman and Adams in the late
1960s – fibrinogen exchange
from various types of surfaces
• Protein B - Initially adsorbed to
the surface
• Displaced in an exchange
reaction by protein A with better
residue functional groups to form
more stable bonds with the
available binding sites on the
surface
• First believed to be unique for
fibrinogen exchange, but it is
general to many proteins
• Influenced by protein properties,
surface properties, protein
concentration, and time
• Proteins with high surface affinity
may not exhibit it
Non-fouling
surfaces
Surfaces that are resistant to
protein adsorption
Represent a special case in
the study of protein–surface
interactions
Development of non-fouling
surfaces is of substantial
interest for marine, food
processing, bioMEMs,
biosensor, and development
of medical devices with
improved biocompatibility
Monolayer vs. Multilayer Adsorption
Key terms

• Surface Modification
• Surfaces can be modified to control protein interactions – Enhance
biocompatibility or reduce fouling
• Techniques used include surface functionalization
• Analytical Techniques
• Radiolabeling
• Fluorescence/surface plasmon resonance
• Quartz Crystal Microbalance
• Materials Selection
• You should select material with appropriate surface properties
Contents of blood

20
SERUM PROTEINS
The protein concentration in plasma/serum is
approximately 60–80 mg/mL of which about 50–60%
are albumins, 40% globulins (10–20%
immunoglobulin G, IgG) and 4% Fibrinogens

Blood protein Normal level %


Function
Albumins 3.5-5.0 g/dl
60% create oncotic pressure 66kD
and transports other molecules

immunoglobulins 1.0-1.5 g/dl 18%


participate in immune 150kD system

Fibrinogens 0.2-0.45 g/dl 4%


blood coagulation 350kD

21
FACTORS AFFECTING PROTEIN
ADSORPTION
• Cell adhesion to artificial materials is affected by their
surface properties such as wettability, roughness, surface
charge, and chemical functionalities
• Wettability: Proteins adsorb more to hydrophobic
surfaces than hydrophilic surfaces
• Roughness: Adhesion is better on roughened surfaces
• Surface charge: Since proteins are charged moles, their
adsorption depends on the law of electrostatics
• Chemical composition: Proteins adhered well to COOH–
and NH2, whereas poorly to CH3– and OH–
• Temperature: The amount of protein adsorbed at high
temperature is usually higher than that at room
22
temperature.
Protein adsorption
measurement
• Radiolabeling
• Fluorescence/surface plasmon resonance
• Quartz Crystal Microbalance
• http://www.youtube.com/watch?v=NyscQUWxGoE

23
Cell attachment, adhesion, spreading,
proliferation, migration
Focal adhesions
• Large, dynamic
protein complexes
through which the
cytoskeleton of a
Focal cell connects to
adhesions the ECM.

Cytoskeleton
• Helps cells
cytoskeleto maintain their
n shape and internal
organization
• Provides
mechanical
support that
enables cells to
carry out essential
functions like
24
division and
Read about these terms

• Cell attachment
• Cell Spreading
• Cell proliferation
• Cell migration

25

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