Chapter 3: Protein Structure and Function Flashcards
Protein is derived from the greek wor ____ meaning ____
proteios means “first”
Why does the term protein derived from the Greek word proteios?
To indicate the central roles that proteins play in the living organisms
the indespensable agents of biological function
Proteins
building blocks of proteins
Amino acids
The stunning diversity of the thousands of proteins found in nature arises from the intrinsic properties of _ _ _ _ _ _
only 20 commonly occurring amino acids
These features include:
- the capacity to polymerize
- novel acid-base properties
- varied structure and chemical functionality in the amino acid side chains
- Chirality
- or handedness
- means that an object or molecule cannot be superimosed on its mirror image by any translation or rotations
Chirality
Classification of proteins
Structural
Contractile/Movement
Transport
Storage/Nutrient
Hormone/Regulatory
Enzyme
Protection/Defense
- are biological catalysts
- majority of this that have been studied are proteins
Enzymes
Function of Enzyme
Catalyze biochemical reactions in the cells
Examples of Enzyme
- Sucrose
- Trypsin
It catalyzes the hydrolysis of sucrose
Surcose
It catalyzes the hydrolysis of proteins
Trypsin
Example of the function of Enzymes
Digestive enzymes pepsin, trypsin, and chymotrypsin break down proteins in our diet so that subunits can be absorbed for use by our cells
Without _ , the body
cannot _ _
- enzymes
- absorb nutrients
- it includes antibodies
- specific protein molecules produced by specialized cells of the immune system in response to foreign antigens
Protection or Defense proteins
Antibodies also called as
Immunoglobulins
These foreign invaders include _ and _ that infect the body
bacteria and viruses
Each _ has regions that precisely fit and bind to a single _
- each antibody
- antigen
Antibody helps to end the ___ by _____ to the ______ and helping to _____ it or ______ it from the body.
- infection
- binding to the antigen
- destroy
- remove
Function of Protection/Defense Protein
Recognize and destroy foreign substances
Example of Protection/Defense Protein
Immunoglobulins
It stimulate immune responses
Immunoglobulins
carry materials from one place to
another in the body
Transport proteins
The protein that transports iron from the liver to the bone marrow
Transferrin
Transferrin is used to
it is used to synthesize the
heme group for hemoglobin
Transferrin is ______ and _______ into ____ mostly by the ___
- synthesized and secreted
- serum
- liver
Synthesis of transferrin is regulated by _____
iron
Iron alone is _______ ______
extremely reactive
If iron is not bound by specific _____ ____ and/or _____ _____ within the body, it can _____ ____ with ____, _____, and _____ structures
- serum carriers
- storage proteins
- viciously interact
- vascular, cellular, and subcellular
Therefore, after absorption, it is bound to the _______ ____ _____ (TF) for ____ _____
- plasma protein transferrin
- safe transport
The proteins that are responsible for transport and storage of oxygen in higher organisms, respectively
Hemoglobin and myoglobin
Function of Transport proteins
carry essential substances throughout the body
Example of Transport proteins
- Hemoglobin
- Lipoproteins
transports oxygen
Hemoglobin
transport lipids
Lipoproteins
Structural Differences of Myoglobin and Hemoglobin:
Myoglobin
- 1 subunit
- heme group
Structural Differences of Myoglobin and Hemoglobin
Hemoglobin
- 4 subunits
- 4 hemegroups
- it controls many aspects of cell function, inlcuding metabolism and reproduction
- can only function within a limited set of conditions
Regulatory/Hormone proteins
Function of Regulatory/Hormone protein
Regulate body metabolism and the nervous system
In regulatory proteins, what must be carefully regulated for life to exist?
- body temperature
- the pH of the blood
- blood glucose levels
Example of Regulatory/Hormone proteins
- Insulin
- Growth hormone
It regulates blood glucose level
Insulin
A hormone that regulate body function (proteins)
Insulin and Glucagon
The hormone that preps your body to cope with stress
Adrenaline: Fight or Flight
How your body creates Adrenaline cells?
- eating high-protein foods,your liver extracts amino acids and sends them to the adrenal glands where they get reshaped into adrenaline and stores
How your body uses Adrenaline cells?
EMERGENCY
- your adrena glands release adrenaline into the blood in a “mass discharge”, your body shifts into “fight or flight” mode
Adrenaline: Fight or Flight
More blood travels to your brain:
Your mind is sharper
Adrenaline: Fight or Flight
Pupils dilate:
Your vision is clearer
Adrenaline: Fight or Flight
The airways in the lungs dilate:
Taking in more oxygen
Adrenaline: Fight or Flight
The hearts contracts more forcefully:
Pumps out more blood
Adrenaline: Fight or Flight
Sweat:
You sweat more
Adrenaline: Fight or Flight
The blood clots more readily:
Helps minimize blood loss
Adrenaline: Fight or Flight
The available source of fuel:
Blood levels of cholesterol, glucose and fatty acids, increases
Adrenaline: Fight or Flight
Skeletal muscles:
More blood gets more oxygen and glucose which strengthen the skeletal muscles
Adrenaline: Fight or Flight
Hunger is no longer a priority:
Blood vessels feeding the gastrointestinal tract narrow and digestive movements slow
How many minutes does these adrenalized effects last?
1 or 2 minutes
What happens to the adrenaline chamiclas after the threat passes?
The adrenaline chemicals oxidize and are converted into waste-product chemicals and are shipped out in urine
It provides mechanical support to large animals and provide them with their outer coverings
Structural proteins
Function of Structural proteins
Provide structural components
Example of Structural proteins
- Collagen
- Keratin
It is in tendons and cartilage
Collagen
It is in hair, skin, wool, and nails
Keratin
Which part of our body that are largely composed of the protein keratin?
Hair and fingernails
What is EB?
Epidermolysis bullosa
Babies that are born with EB are known as?
“butterfly babies”
People with EB have __ ___ resulting to ___ ____ ___ in the skin
- genetic mutation
- abnormal structural proteins
What is the effect of recessive Dystrophic EB?
- difficulty to ingest food for a child
- causes blistering that occurs in mouth, esophagus, and gastrointestinal tract
It is necessary for all forms of movement
Movement/Contractile proteins
Function of movement/contractile protein
Make muscles move, including the heart
Example of Movement/Contractile protein
- Myosin
- Actin
It contracts muscle fibers
Myosin and actin
It serve as sources of amino acids for embryos or infants
Nutrient proteins
Examples of Storage/Nutrient protein
- Casein
- Ferritin
- Egg albumen
It stores protein in milk
Casein
It stores iron in the spleen and liver
Ferritin
these compounds contain both an amine and an acid
Amino acids
20 amino acids that are common in nature are all?
All 20 are α-amino acids
α (alpha) means
amine is adjacent to the carboxylate group
Out of all 20 amino acids, how many are stereoisomers?
19
What amino acid is not a stereoisomer out of all 20 common amino acid?
Glycine (does not have a chiral carbon)
The α-carbon of amino acids is ____
Chiral
What is a chiral molecule?
It is non-superposable to its mirror image due to the presence of an asymmetric carbon atom
Where does all the differences between amino acids depens upon?
side-chain R groups
Forming classes of Amino acids is based on, what?
Polarity of their side chains
Classes of Amino Acids
- Nonpolar
- Polar, neutral
- Polar acidic
- Polar basic
It is a class of amino acid that has hydrophobic R groups
Nonpolar
It is a class of amino acid that have a high affinity for water, but are not ionic at pH
Polar, neutral
- it is a class of amino acid that have ionized carboxyl groups in their side chains
- negatively charged
Polar acidic
- are basic as the side chain reacts with water to release a hydroxide anion
- positively charged
Polar basic
What are the amino acids that are essential for normal tissue growth and development?
All amino acids
What is the term that is reserved for those amino acids that must be supplied in the diet for proper growth and development?
“essential amino acids”
“PVT. TIM HALL”
Phe, Val, Thr, Trp, Ile, Met, His, Arg, Leu, Lys
___ and ___ are semi-essential; they not synthesized in sufficient quantities during infancy stage
His and Arg
α-carbon is attached to a:
- Carboxyl group (̶ COOH)
- Amino group ( ̶ NH2)
At physiologic pH the amino acid has:
- Carboxyl group in –COO-
- Amino group in –NH3+
- Neutral molecule with equal number of + and – charges is called a zwitterion
What is a zwitterion?
Neutral molecule with equal number of + and – charges
From the German word “zwitter” which means
hybrid or hermaphrodite
Amino acids are __ ____ __ with __ ___ __ and __ ___ ___
- white crystalline solids
- high melting points
- high water solubilities
What are the two charged groups?
- basic amino group
- carboxylic acid
The two charged groups at the two ends lead to
internal proton transfer, forming zwitterions
By changing the __ you can affect the __ ___ on the zwitterions
- pH
- net charge
It is the pH point at which there is no net charge on the zwitterions
Isoelectric point (pI)
What is the isoelectric point of an amino acid?
the pH at which it bears no net charge
What is a form of acid that is more accurate depiction of its structure?
Zwitterionic
A ____ contains two point charges but is ___ overall
- zwitterion
- neutral
At pH values below isoelectric point
bear net positive charge
At pH above isoelectric point
bear net negative charge
It is a test by applying a sample of the amino acid to specially treated paper or gel and applying an electric field at different pH values
Electrophoresis
A molecule with a net charge of zero will __ migrate in an electric field
not
positive charge (+) will migrate toward
cathode (-)
negative charge (-) will migrate toward
anode (+)
For a typical amino acid with a neutral sidechain R:
- the positively charged form (+1) dominates at low pH
- the zwitterionic (neutral) form dominates at intermedaite pH
- the negatively charged form (-1) dominates at high pH
For an amino acid with an acidic sidechain:
- the positively chargde form (+1) dominates very low pH
- the zwitterionic (neutral) form dominates at low pH
- the negatively charged form (-1) dominates at intermediate pH
- at strong basic pH a doubly negatively charged form (-2) may form
For an amino acid with a basic sidechain (e.g. lysine)
- the doubly positive charged form (+2) dominates at very low pH
- the positively charged form (+1) dominates at intermediate pH
- the zwitterionic form (neutral) forms at basic pH
- at strongly basic pH, the negatively charged form (-1) dominates
What is the pI values of the polar acidic amino acids?
pH 3
What are the polar acidic amino acids?
- Aspartic acid
- Glutamic acid
The carboxylic acid in R groups of their zwitterions at pH 3 is __ ___
Not ionized
The carboxylic acid in R groups at physiological pH __ _ to form a ___ ___ `
- loses H+
- negatively charged
What is the pI values of basic amino acids?`
typically higher than physiological pH value, ranging from pH 7.6 to 10.8
The amines in the R groups of the basic amino acids at physiological pH values __ _ to form an ___ ___ __
- gain H+
- overall positive charge
What are the basic amino acids?
- lysine
- arginine
- histidine
Amino acid:
Neutral
Isoelectric pH:
4.8 - 6.3
Example of Neutal Amino Acid
Ala (6.1)
Amino acid:
Acidic
Isoelectric pH:
2.8 - 3.2
Example of Acidic Amino Acid
Asp (2.8)
Amino acid:
Basic
Isoelectric pH:
7.8 - 10.8
Example of Basic Amino Acid
Lys (9.7)
It is an analytical method for identifying amino acids by observing their migration as a function of pH under an applied electric field gradient
Electrophoresis
If an amino acid carries a positive charge at pH<pI, it will ___
migrate to the negative electrode (cathode)
If an amino acid carries a negative charge at pH>pI, it will ___
migrate to the positive charge electrode (anode)
What are the techniques in separating amino acids?
- Ion-exchange chromatography
- Size-exclusion chromatography
- Affinity chromatography
Ion-excahnge chromatography
Separates the compounds base on charge
Size-exclusion chromatography
Separates compounds base on size
Affinity chromatography
Separates the compounds base on the interaction between the protein of interest and ligand
A result of covalent bonding between the amino acids
The Peptide Bonds
In peptide bonds, proteins are?
linear polymers of L-α-amino acids
The peptide bond:
Carboxyl group of one amino acid is linked to ___
the amino group of another amino acid
It is an amide bond or peptide linkage
Linkage
What is the reaction involve in peptide bond?
Dehydration reaction
What is produce if the two amino acids are condensed or dehydrated?
Dipeptide
N-terminal amino acid
Amino acid with a free a-NH3+ group
C-terminal amino acid
Amino acid with a free –COO ̶ group
Amino acid structures are written with the ___
N-terminal on the left
amino acids are polymerized into ___ and ___
peptides and proteins
It is used for molecules composed over 50 amino acids
Protein
It is used for molecules of less than 50 amino acids
Peptide
Naming Peptides
- The far right AA residue retains the name of the amino acid
- All AAs (except TRYPTOPHAN) → - ine or –ic acid is replaced by –yl
- Tryptophan becomes tryptophanyl
Small Peptides
- Aspartame (Asp-Phe)
- Glutathione (Glu-Cys-Gly)
- Enkephalins (Tyr-Gly-Gly-Phe-Leu & Tyr-Gly-Gly-Phe-Met)
- Oxytocin (Ile-Leu)
- Vasopressin (Phe-Arg)
- it is the artificial sweetener used in almost every diet food on the market today
- its caloric content is the same as sucrose but is ~180 times as sweet
Aspartame (Asp-Phe)
Forms have a bitter taste
- L-D
- D-L
- D-D
- it functions as an antioxidant, protecting cellular contents from oxidizing agents such as peroxides and superoxides
Glutathione (Glu-Cys-Gly)
What are the oxidizing agents that are highly reactive forms of oxygen often generated within a cell?
Peroxides and Superoxides
Glu is bonded to Cys through the ___ ___ ___ rather than through the ___ ____ ___
- side-chain carboxyl group
- α-carbon carboxyl group
- natural painkillers produced in the body
- it is bind to receptors in the brain to give relief from pain
Enkephalins and Endorphins
- it is the first hormone to be synthesized in the laboratory
- a nonpeptide used to initiate labor
Oxytocin
Oxytocin stimulates ____ _____ in labor
uterine contractions
- an antidiuretic hormone
- regulates blood pressure by adjusting the amount of water reabsorbed by the kidneys
Vasopressin
- partly responsible for triggering pain, welt formation (as in scratches), movement of smooth muscle, and lowering of blood pressure
Bradykinin (Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg)
completely inactive, hence, the name bogus or false
Boguskinin (Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe- )
Val-His-Leu-Thr-Pro-Glu-Glu-Lys-Ser-Ala-
Normal Hb
Val-His-Leu-Thr-Pro-Val-Glu-Lys-Ser-Ala-
Sickle-cell Hb
Val-His-Leu-Thr-Pro-Glu-Lys-Lys-Ser-Ala-
Georgetown anemia
The sickled cells are unable to pass through the ___ _____ of the circulatory system, and circulation is ____
- small capilliaries
- hindered
Effect of sickled cells in human body
- damage to many organs (bone and kidney)
- can lead to death at an early age
It is results from formation of covalent peptide bonds between amino acids
Primary Structure of Proteins
What is the Primary Structure of Proteins?
The amino acid sequence of the polypeptide chain
The peptide bond has a __ ___ __ character
partial double bond
There is __ ___ around __ __ of the three single bonds of a peptide backbone
- free rotation
- only two
R groups on adjacent amino acids are on ____ ___ of the chain because of the rigid peptide bond.
Opposite sides
How does the Second Structure of Proteins is formed?
When the primary sequence of the polypeptide folds into regularly repeating structures
Secondary structure results from ___ ___ between the _____ ___ and ______ ___ of the peptide bonds
- hydrogen bonding
- amide hydrogens (N—H)
- carbonyl oxygens (C=O)
Most common type of secondary structure
- Coiled
- Helical
Secondary Sructure of Proteins includes:
α-helix and β-sheet
Important features of Secondary Structure of Proteins: α-Helix
- Each amide H and carbonyl O is involved in H bonds locking the helix in place
- Carbonyl O links to amide H 4 amino acids away
- H bonds are parallel to the long axis of the helix
- Helix is right-handed
- Repeat distance or pitch is 5.4 angstroms
- 3.6 amino acids per turn
In α-Helix, the borken line represents the? (tanawa lng nnyo sa pdf na gi send ni maam ang picture hehe sankss)
Hydrogen bonds
What is the direction Hydrogen bonds in α-Helix?
Parallel to the direction of the helix
are arranged in a secondary structure of fibers or sheets with only 1 type of secondary structure
Fibrous proteins
- it is the second most common secondary structure
- appears similar to folds of fabric
β-pleated sheet
In β-pleated sheet, what organic compounds that are involved in the H bonds with the chain nearly completely extended?
- carbonyl O
- amide H
β-pleated sheet:
Two possible orientation
- Parallel
- Antiparallel
Two possible orientation:
Parallel
The N-termini are head-to-head
Two possible orientation:
Antiparallel
The N-terminus of one chain is aligned with the C-terminus of the other
- three-dimensional structure
- overall folding of the entire polypeptide chain
- defines the biological function of proteins
The Tertiary Structure of Proteins
- forms spontaneously
- maintained by interactions among the side chains or R groups
Globular tertiary structure
Types of Interactions Maintaining Tertiary Structure
- Disulfide bridges
- Salt bridges
- Hydrogen bonds
- Hydrophobic interactions
Disulfide bridges
interaction between two cysteine residue
Linkage that occurs within the same chain
Intrachain
Linkage the occurs between 2 or more chains
Interchain
Salt bridges
- ionic interaction/electrostatic attraction
- interaction between ionic side chains –COO– and –NH3+
Hydrogen bonds
Interaction between polar residue side chains
Hydrophobic interactions
two nonpolar groups are attracted by a mutual repulsion of water
- it is the arrangement of subunits or peptides that form a larger protein
- is maintained by the same forces which are active in maintaining tertiary structure
The Quaternary Structure of Proteins
What is the functional form of many proteins?
Aggregate of several globular peptides
A polypeptide chain having primary, secondary, and tertiary structural features that is a part of a larger protein
Subunit
Protein Functions Follow Shape
Fibrous proteins:
- Mechanical strength
- Structural components
- Movement
Protein Functions Follow Shape
Globular proteins:
- Transport
- Regulatory
- Enzymes
Shape of Globular proteins
Roughly circular
Shape of Fibrious proteins
Long strands
Amino acid sequence of Globular proteins
Irregular and wide range of R groups
Amino acid sequence of Fibrious proteins
Repetitive with a limited range of R groups
Function of Globular proteins
Physiological/functional
Function of Fibrious proteins
Structural
Examples of Globular proteins
- Hemoglobin
- Enzymes
- Insulin
- Immunoglobulin
Examples of Fibrous proteins
- Collagen
- Keratin
- Myosin
- Actin
- Fibrin
Solubility of Globular proteins
(generally) soluble in water
Solubility of Fibrous proteins
(generally) insoluble in water
Hemoglobin in Normal (wild-type)
Normal (Globular)
Red Blood Cell in Normal (wild-type) Hemoglobin
Round (biconcave)
Blood Vessels in Normal (wild-type) Haemoglobin
Free flowing
Hemoglobin in ‘Sickle Cell’
Clumped (fibrous)
Red Blood Cell in ‘Sickle Cell’ Hemoglobin
Sickle-shaped
Blood Vessels in ‘Sickle Cell’ Hemoglobin
Forms clots/blockages
It is a protein to which another chemical group (e.g., carbohydrate) is attached by either covalent bonding or other interactions
Conjugated Proteins
It is the oxygen storage protein of skeletal muscle
Myoglobin
It is the oxygen-transport protein of higher animals
Hemoglobin
Why does the oxygen is transferred from hemoglobin to myoglobin?
Since the myoglobin has a stronger attraction for oxygen than hemoglobin does
It is an essential component of the proteins hemoglobin and myoglobin
Heme group
In the heme group, it is the oxygen binding site
Fe2+, ferous ion
Each hemoglobin contains a ___ ___ which can hold __ ____ of oxygen (O2)
- heme group
- 1 molecule
Hemoglobon
T-form:
- taut or tense
- Deoxy: low O2 affinity
Hemoglobon
R-form:
- relaxed
- Oxy: high O2 affinity
The disruption of bonds in the secondary, tertiary, and quaternary protein structures
Protein Denaturation
It is the loss of organized structure of a globular protein
Denaturation
It is the breaking of bonds using water
Hydrolysis
- splits the peptide bonds to give smaller peptides and amino acids occurs in the digestion of proteins
- occurs in cells when amino acids are needed to synthesize new proteins and repair tissues
Protein Hydrolysis
In the lab, the hydrolysis of a peptide requires __ or ___, ___, and ___
acid or base, water, and heat
In the body, _____ catalyze the hydrolysis of proteins
Enzymes
Some practical aspects of protein denaturation
- Heat and UV
- Salts of heavy metal ions esp. Hg2+, Pb2+, Ag+
- Organic compounds such as soap, detergents, phenol, and aliphatic alcohol
Protein Sequencing
Steps in determining the amino acid sequence:
- hydrolysis - by acid, alkali, or enzyme
- identification of the products of hydrolysis
- fitting the pieces together as you would a jigsaw puzzle
Protein Sequencing
HYDROLYSIS
- Acid Hydrolysis
- Alkali Hydrolysis
- Cyanogen bromide
- Enzymatic Hydrolysis by proteases/ peptidases
- involves heating in the presence of 6N HCl
- the protein is completely hydrolyzed, but Trp, is destroyed completely and Ser, Thr, and Tyr are partially destroyed
Acid Hydrolysis
- heating in the presence of 4N NaOH
- does not damage Trp, but destroys Arg, Cys, Thr, & Ser; and some amino acids are partly deaminated
- more disadvantageous
- it is used in quantitative determination of this amino acid
Alkali Hydrolysis
- cuts peptide bonds on the carboxylterminal side of methionine residues
- used to reduce the size of polypeptide segments for identification and sequencing
Cyanogen bromide
Two types of Enzymatic Hydrolysis
- Exopeptidases
- Endopeptidases
Enzymes that cleave external peptide bonds
Exopeptidases
Exopeptidases is used in __
the determination of the amino
acid sequence of peptides
Enzymes under Exopeptidases
- Aminopeptidases
- Carboxypeptidases
It sequentially cleaves peptide bonds, beginning at the N-terminal end of the polypeptide
Aminopeptidases
It sequentially cleaves peptide bonds beginning at the C-terminal end of the polypeptide
Carboxypeptidases
Enzymes that cleave internal peptide bonds
Endopeptidases
Enzymes under Endopeptidases
- Trypsin
- Chymotrypsin
- Elastase
- Pepsin
- Thermolysin
It cleaves peptide bonds at the carboxyl end of the two strongly basic amino acids: arginine and lysine
Trypsin
It cleaves peptide bonds at the carboxyl end of the three aromatic amino acids: phenylalanine, tyrosine, & trptophan; and Leucine
Chymotrypsin
It cleaves on the carboxyl side of Gly and Ala
Elastase
It cleaves peptide bonds at the amino end of the three aromatic amino acids: phenylalanine, tyrosine, tryptophan; acidic amino acids, Asp and Glu; and Ile
Pepsin
It cleaves peptide bonds at the amino end of the three aromatic amino acids, Phe, Tyr, Trp; and amino acids with bulky nonpolar R groups, Leu, Ile, and Val
Thermolysin
Other methods for determining the N-terminal end (chemical method)
- Sanger’s Method
- Edman Degradation
The key reagent in this method for identifying the N-terminus is 1-fluoro-2,4-dinitrobenzene
Sanger’s Method
- can be done sequentially one residue at a time on the same sample
- usually, one can determine the first 20 or so amino acids from the N-terminus by this method
Edman Degradation
What is the key reagent in the
Edman degradation?
phenyl isothiocyanate
Other methods for determining the C-terminal end (chemical method)
Hydrazine Method
Hydrazine reacts with all amino acids whose carboxyl group is bound in peptide linkage, creating ____ ____ ____
Amino acyl hydrazides
It regulates body growth
Growth hormones