Biology Hidden Gems Must Memorize Flashcards
Fatty Acids
Building blocks for most but not all complex lipids.
Long chains of carbon truncated on one end by COOH.
Can be Saturated or Unsaturated (by hydrogens)
Triacylglycerols
Store Metabolic energy, provide insulation and padding.
Sometimes called triglycerides or simple fats and oils
Constructed from a three carbon backbone named glycerol
Adipocytes
Fat cells that are specialized cells where their cytoplasm contains nothing but triglycerides
Phospholipids
Serve as a constructional component of membranes.
Built from a glycerol backbone but a polar phosphate group replaces one of the fatty acids.
Amphipathic-have two charges on two different sides of the molecule.
Steroids
Four ringed structures including hormones, vitamin D, and cholesterol (a vital membrane component)
Regulate metabolic activities
Lipoproteins
Transfer lipids that are insoluble in aqueous solution
Contains a lipid core surrounded by phospholipids and apoproteins
Classified by their density. VLDL, LDL, HDL
Proteins (Polypeptides)
Built from a chain of amino acids held together by peptide bonds
Important Amino Acids
Proline-disrupts alpha-helices
Alanine-a methyl group
Glycine-just a hydrogen
Cysteine/Methionine- both contain a sulfur
Both alpha helices and beta sheets reinforced by hydrogen bonds between the carbonyl oxygen and the hydrogen on the amino group.
Five Forces Create tertiary structure
Covalent disulfide bonds between two cysteine amino acids on different parts of the chain.
Electrostatic (ionic bonds): Mostly between acidic and basic side chains
Hydrogen Bonds
Van der Wals forces
Hydrophobic side chains pushed away from water towards the center of the protein.
When you see Nitrogen THINK…
Protein
Glycoproteins
Proteins with carbohydrate groups attached
Proteoglycans
Mixture of proteins and carbohydrates
Anomers
-Alpha- When the hydroxyl group on the first carbon and the methoxy group on the 6-carbon are on opposite sides.
Glycogen
- Found in all animal cells, large amounts found in muscle and liver.
- Liver regulates blood glucose levels so they are one of the few cell types capable of reforming glucose from glycogen and releasing it back into the blood stream.
- Only certain epithelial cells in the digestive tract and the proximal tubule of the kidney are capable of absorbing glucose against a concentration gradient. This is done via secondary transport down the sodium concentration gradient.
Insulin
Increases the rate of facilitated diffusion for glucose and other monosaccharides.
In the absence of insulin only neural and hepatic cells are capable of absorbing sufficient amounts of glucose via the facilitated transport system.
Minerals
DIssolved inorganic ions inside and outside the cell. By creating electrochemical gradients they assist in the transport of stuff into and out of the cell.
Enzyme Models
Lock and Key- the active site of the enzyme has a specific shape that only binds the specific substrate
Induced fit model-The shape of both the enzyme and the substrate are altered upon binding
Enzyme Kinematics
Vmax is proportional to enzyme concentration
Km does not vary with enzyme concentration and therefore is a good indicator of an enzyme’s affinity for its substrate
Cofactor
non-protein component required by an enzyme to reach the optimal activity; either mineral or coenzyme
Apoenzyme
An enzyme without its cofactor
Coenzyme
Cosubstrates and prosthetic groups
Cosubstrates
Reversibly bind to a specific enzyme and transfer some chemical group to another substrate. The cosubstrate is then changed back to it’s original form by another enzymatic reaction
Prosthetic groups
Remain covalently bound to the enzyme throughout the reaction
Competitive inhibition
Raise the apparent Km but do not change the Vmax
Can be overcome by excess substrate
Noncompetitive inhibition
Can’t be overcome by excess substrate
Vmax is lowered but since enzyme affinity is the same Km remains the same
Vmax
The rate of reaction when the enzyme is saturated with substrate is the maximum rate of reaction,
Vmax is the reaction rate when the enzyme is fully saturated by substrate, indicating that all the binding sites are being constantly reoccupied
Km
Km is the concentration of substrate which permits the enzyme to achieve half Vmax. An enzyme with a high Km has a low affinity for its substrate, and requires a greater concentration of substrate to achieve Vmax
Uncompetitive Inhibition
Uncompetitive inhibition, also known as anti-competitive inhibition, takes place when an enzyme inhibitor binds only to the complex formed between the enzyme and the substrate (the E-S complex). Uncompetitive inhibition typically occurs in reactions with two or more substrates or products.
Km reduced and Vmax reduced
Line weaver Burke plots for inhibition
Feedback inhibitors do not resemble…
The substrate of the enzyme they inhibit
Allosteric regulation
Come back and bind to a different site of the enzyme causing a conformational change, many alter affinity of substrate for enzyme (Km) but not Vmax.
Can be either positive or negative regulation
At low substrate concentrations, small increases in substrate concentration increase enzyme efficiency as well as reaction rate.
Lyase
Catalyzes the addition of one substrate to a double bond while a ligase governs an addition reaction using ATP.
Metabolism
1) Macromolecules are broken down into their constituent parts releasing little or no energy
2) Constituent parts are oxidized to acetyl CoA, pyruvate or other metabolites forming ATP and reduced enzymes (NADH and FADH2)
3) If oxygen is available and the cell is capable of using oxygen these metabolites go into the citric acid cycle and oxidative phosphorylation to form large amounts of energy (more NADH, FADH2, or ATP); otherwise the coenzyme NAD+ and other byproducts are either recycled or expelled as waste.
Glycolysis
First step of anaerobic and aerobic respiration
- Starting molecule is glucose
- Products: 2 molecules of ATP from ADP/water, 2 pyruvate molecules from 1 glucose, 2 molecules of NADH from reduction of NAD+
- In the third step the molecule is committed to glycolysis
- Regulated and committed steps are 1,3,10. Very - delta G. Very spontaneous
Substrate level phosphorylation
The formation of ATP from ADP using energy from the decay of high energy phosphorylated compounds as opposed to using energy from diffusion (oxidative phosphorylation)
-2 ATPs are spent, 4 ATPs are produced
Much of the fructose and galactose ingested by humans is converted into glucose in the liver enterocytes; however fructose can enter as
fructose-6-phosphate or G3P; galactose can be converted to G6P to enter glycolysis.
Fermentation
- Anaerobic respiration
- Includes glycolysis, reduction of pyruvate to ethanol or lactic acid, and oxidation of NADH back to NAD+
- Takes place when a cell or organism is either unstable to assimilate the energy from NADH and pyruvate, or has no oxygen available to do so
- Recycles NADH back to NAD+
Aerobic Respiration
- Products of glycolysis move into the matric of the mitochondrion
- Once inside the matrix, pyruvate is converted to acetyl CoA in a reaction that produces NADH and CO2
Krebs Cycle
- Acetyl CoA transfer two carbons from pyruvate to 4-carbon oxaloacetic acid
- Each turn produces 1 ATP, 3 NADH, and 1 FADH2
- ATP production in the Krebs cycle is substrate-level phosphorylation
- Triglycerides can also be catabolized for ATP. Fatty acids are converted into acyl CoA along the outer membrane of the mitochondrion and endoplasmic reticulum at the expense of 1 ATP.
- The reaction also produces FADH2 and NADH for every two carbons taken from the original fatty acid
- The glycerol backbone is converted to PGAL
Amino acids are deaminated by…
the liver. The deaminated product is either chemically converted to pyruvic acid or acetyl CoA, or it may enter the Krebs cycle at various stages depending upon which amino acid was deaminated.
Electron Transport Chain
- A series of proteins on the inner membrane of the mitochondrion
- The first protein complex in the series oxidizes NADH by accepting high energy electrons that it will then pass to O2
- As electrons are passed along, protons are pumped into the intermembrane space for each NADH
- The protons then diffuse back to the mitochondrial matrix turning ADP into ATP through the pump, ATP synthase
Oxidative phosphorylation-production of ATP through diffusion/oxidation of NADH, like this
Intermembrane space has lower pH than matrix. (More acidic because of all the protons being pumped out into it.
Aerobic respiration produces how many net ATPs?
About 36 net
How many ATP does 1 NADH bring back?
2 to 3 ATPs
How many ATP does 1 FADH2 bring back?
2 ATPs
Glucose + O2—–>
CO2+H2O
Not balanced here, its a combustion reaction
Gene
Series of nucleotides that code for the production of a single polypeptide or mRNA, rRNA, tRNA
Eukaryotes can have multiple copies or a gene but prokaryotes only have one copy of each gene.
Eukaryotic genes that are actively transcribed by a cell area associated with regions called
Euchromatin
Genes not being actively transcribed are associated with tightly packed regions called
Heterochromatin
Genome
Entire sequence of DNA of an organism
Central Dogma
DNA is transcribed to RNA, which is translated to amino acids forming a protein
DNA
Sugar phosphate backbone
Phosphodiester bond
3’ end attached to an -OH group and 5’ end attached to a phosphate group
A2T hydrogen bonds, C3G hydrogen bonds
Semi Conservative DNA replication
When a new strand is created one strand from the original DNA goes to the new strand.
What is DNA replication governed by?
Replisome.
Two replisomes proceed in opposite directions along the chromosome making replication a bidirectional process.
The point where a replisome is attached to the chromosome is called the replication fork.
Replicons
Replication units- each chromosome of eukaryotic DNA is replicated in many discrete segments called replicons.
DNA helicase
Unwinds double helix
DNA polymerase
Enzyme that builds the new DNA strand, can’t initiate a strand from two nucleotides, needs primer
Reads from 3’—> 5’ (upstream direction), writes 5’—>3’ (downstream)
Primase
-an RNA polymerase creates an RNA primer 10 ribonucleotides long to initiate the strand
Lagging strand
Interrupted strand
Made up of Okazaki Fragments
Leading Strand
Written Continuously
Since the formation of one strand is continuous and the other fragmented, the process of replication is called semi discontinuous
DNA ligase
Moves along lagging strand and ties the Okazaki fragments together
Exonuclease
Removes nucleotides from the center of strand, exonuclease on DNA polymerase
Telomeres
Repeated six nucleotide units that protect the chromosomes from being eroded through repeated rounds of replication
Telomerase
Catalyzes the lengthening of telomeres
Differences between DNA and RNA
RNA is produced by transcription-RNA is manufactured from a DNA template in this process
-DNA is produced by replication
rRNA (ribosomal RNA)
Combines with proteins to form ribosomes
Initiation
Beginning of transcription
- An initiation factor finds a promoter on the DNA strand
- Promoter- sequence of DNA nucleotides that designates a beginning point for transcription
In prokaryotes its located upstream
Most commonly found sequence is the consensus sequence
Variations from the consensus sequence causes RNA polymerase to bond less tightly and less often leading to those genes being transcribed less often.
After binding to the promoter RNA polymerase unzips the double helix creating a transcription bubble, next the complex switches to elongation mode
Elongation
RNA polymerase transcribes only one strand or antisense strand.
The other strand, the sense strand (coding strand) protects its partner against degradation
RNA polymerase moves along the DNA strand from 3’—>5’ building the new RNA 5’—>3’. NO proofreading, 10X slower than replication
Termination
Requires special termination sequence to tell RNA polymerase to detach.
- Replication makes no distinction between genes, transcription does through activators and repressors
- Most genetic regulation occurs at transcription when regulatory proteins bind DNA and activate or inactivate its transcription.
Activators and Repressors
Bind to DNA close to the promoter and either activate or repress the activity of RNA polymerase (activators before the promoter sequence, repressor after)
- Primary function of gene regulation in prokaryotes is to respond to environmental changes
- Changes in gene activity are a response to the concentration of specific nutrients in and around the cell
- Primary function of gene regulation in multicellular organisms is to control intracellular and extracellular environments
Polycistronic
Prokaryotic mRNA typically includes several genes in a single transcript
Monocistronic
Eukaryotic mRNA includes one gene per transcript
Operon
Genetic unit of prokaryotic DNA consisting of the operator, promoter and all other genes that contribute to a single mRNA
Post-transcriptional Processing
Occurs in both eukaryotes and prokaryotes
-In prokaryotes rRNA and tRNA go through post-transcriptional processing but mRNA usually doesn’t
in eukaryotes mRNA goes through post-transcriptional processing too
Primary Transcript
initial mRNA nucleotide, also known as pre-mRNA
- Processed in three ways
1. Addition of nucleotides
2. Deletion of nucleotides
3. Modification of nitrogenous bases
In eukaryotic mRNA the 5’ cap is added as protection against degradation by exonucleases
The 3’ end is polyadenylated with a poly A tail to protect it from exonucleases
Introns
Before leaving the nucleus, introns are removed from the pre-mRNA
Do not code for protein and are degraded in the nucleus
Exons
Parts of pre-mRNA that survive post-transcriptional processing
-can be spliced together in different ways to code for different proteins
snRPs
Recognize nucleotide sequences at the ends of introns
- Several snRPs form a complex called a spliceosome
- Spliceosome-inside introns are looped bringing exons together
Denatured
When heated or immersed in high concentration salt solution or high pH solution
Melting temperature Tm is higher for G-C because they make more hydrogen bonds than A-T
Denatured DNA is less viscous, denser, and more able to absorb UV light
Nucleic Acid Hybridization
Separated strands like to spontaneously associate with their original parter: DNA-DNA, DNA-RNA, RNA-RNA
Restriction enzymes (endonucleases)
Digest (cut) nucleic acid sequences
bacteria defend themselves from viruses by cutting the viral DNA into fragments
Bacteria protect their own DNA with methylation (adding -CH3)
Typically a restriction site is palindromic
Recombinant DNA
DNA that is reformed from restriction endonuclease cuts (remember sticky ends)
Reconnected by DNA Ligase
Can be made long enough for bacteria to replicate and then placed within the bacteria using a vector, typically a plasmid or sometimes a virus
-The bacteria can then begin to be grown in large quantity forming a clone of cells containing the vector with the recombinant DNA. The clones can be saved in a library.
By including in the vector a gene for resistance to a certain antibiotic and the lacZ gene which enable the bacteria to metabolize the sugar X-Gal
When you apply the antibiotic those without the vector will die
You use an endonuclease that cuts at the lacZ gene to insert the new DNA so if the bacteria metabolizes X-Gal you know its not the right bacteria. Clones with the active lacZ gene turn blue.
Complementary DNA (cDNA)
DNA reverse transcribed from mRNA
-It’s useful to clone DNA with no introns so you use reverse transcriptase
Polymerase Chain Reaction
The double strand to be cloned(amplified) is placed in a mixture with primers, polymeraseHeated to 95°C to denature it, cooled to 60°C, the primers hybridize(anneal) to their complementary ends of the DNA strands–Heat resistant polymerase is added and is activated when the temperature hits 72°C
Southern Blotting
- technique to identify target fragments of known DNA sequence in large populations
- DNA is cleaved into restriction fragments which are then resolved by gel electrophoresis
- Large fragments move slower than small ones
Next, the gel is made alkaline to denature the DNA and a sheet of nitrocellulose is used to blot the gel which transfers the resolved single stranded DNA fragments onto the membrane
-A radio labeled probe with a nucleotide sequence complementary to the target fragment hybridizes with and marks the target fragment…this reveals the location of the probe and the target fragment.
Northern Blotting
Just like southern blotting except it identifies RNA