Cell BIO

NOTES:

  • There are higher concentrations of the following in the cell:

    • K+ , Mg+ , PO4+3 , O2

  • There are higher concentrations of the following outside the cells:

    • Na+ , Cl- , HCO-3 , O2

  • Almost all diseases happen because the body is thrown out of homeostasis 

    • “Everything exists to maintain homeostasis”

  • Positive feedback loops are part of a larger negative feedback loop

  • Cells broken up into percentage are made up of the following:

    • 70-85% Water

    • 10-12% Proteins

    • 2% lipids

    • 1-6% carbohydrates

      • Glycogen is higher inside the cell instead of glucose to keep a gradient to be able to pull glucose into a cell.

  • Plasma Membrane:

    • Made up of phospholipids and proteins

      • Phosopholipids are made up of three parts

        • Hydrophilic head

        • Glycerol backbone

        • Lipophilic tail 

    • Cholesterol aids in helping stiffening the membrane

      • The number of cholesterol in the membrane is temperature dependant

  • Membrane proteins have different purposes:

    • Cell adhesion 

      • GPI anchors

    • Transmembrane movement of water soluble substances

      • Pores/chanels 

      • Carriers

      • Pumps 

        • These use ATP

    • Can be enzymes

    • Cell to Cell communication

    • Molecule recognition 

    • Attachment to cytoskeleton

  • Glycocalyx:

    • Membrane proteins with carbohydrate modifications

    • Have different purposes:

      • Negatively charge to repel other negatively charged protiens

      • They can interact with other glycocalyx of other cells

      • Act as receptors (insulin)

      • Important to immune system

  • Cytoskeleton:

    • Provides structural support 

    • Allows cells to maintain shape

    • Maintains Polarity

    • Organizes intracellular organelles

    • Helps with movement and cell migration

    • Helps with transport

      • Microtubules

        • Largest of the filaments

        • Think of them like a hollow straw

        • Made up of a- and b- tubulin dimers 

        • Constant growth and dissolution

        • Polymerization starts at the centrioles in the microtubule organizing center (MTOC)

        • They are the ones that pull the chromosomes to the side during splitting 

        • Found in cilia, mitotic spindles, flagella, and the centrioles “conveyor belt”

          • Cilia is found in lung cells and the ear

          • Flagella is found on sperm cells

          • 9+2 structures

        • They are paths for motor proteins 

        • Kinesin and Dynein

      • Microfilament

        • Smallest of all the filaments but strongest

        • Provides elastic support

        • They attach to the adherin junctions

        • Important in muscle contraction 

          • Actin and myosin

        • Associated with thick filaments (myosin in muscles)

        • Treadmilling constant building and breaking down

        • They consist of a head and a tail ATP hydrolysis causes a pivoting head.

      • Intermediate Filament

        • Think of them as box spring

        • They are permanent stable structure 

        • They connect cellular structures 

        • They are made of two polypeptide dimers wrapped around each other in antiparallel fashion. (rope like structure)

        • Connect desmosomes and hemidesmosomes together

        • Use different bases to make themselves depending on the cell type

          • Kertin (Hair)

          • Lamins

          • Vimentin

          • GFAP

          • Neurofilaments

          • Desmin

          • Cytokeratins

  • Cell Polarity each part of the cell has different job

    • Microvilli not on all sides of the GI cells

    • Directional transport

    • Barrier integrity 

    • Organization

  • Claudins and Occludins means Closed

  • Tight Junctions (Zona Occludens)

    • Water tight “quilting”

    • Formed from alternating seams of claudin and Occludin proteins

    • Barriers: Seal the cells together so ions can't go in between the cells

    • Gates: Can be selective and let certain things in.

    • Fences: Separated the top of the cell and bottom of the cells

  • Anchoring Junctions 

    • Mechanically attach cells together. NEED cadherins to mediate these attachments.

    • Different types:

      • Adherens junctions

        • Connect F-actin filament to cadherin

        • Cadherins form stable cell to cell junctions that link to the cytoskeleton of the other cell

        • Think of them like a belt goes all the way around the cell

        • Influence cell shape and mobility

          • If they don't work in cancer cells that cancer can metastasize 

      • Desmosomes

        • Think of them as “rivets” or “spot welds”

        • Connect the intermediate filament to cadherins

        • Communicate extracellular forces to multiple cells

        • Allows some movement and some water and ions to move in the space between cells

        • Desmosomal cadherins link adjoining cells to intermediate filaments

      • Hemidesmosomes

        • Think of them as “wall screws” or “wall anchors”

        • Transmembrane connectors (integrins) attach to the intermediate filaments to the ECM.

  • Gap Junctions

    • Dense array of pores 

    • “Tunnels” allow exchange of ions, sugars, amino acids, small molecules.

    • Permeability depends on intracellular pH and Ca+

    • Heart has a ton of gap junctions to help iron flow so it can contract unisonly

    • “Communicating junktion”

  •  Mitochondria

    • Power house of the cell

    • Enslaved bacteria (theory)

      • 2 membranes

      • The folds of the inner membrane are called Cristae

    • 2 distinct compartments 

      • Intermembrane space 

      • Matrix space

    • ATP production 

    • Mitochondria genome codes for mitochondrial tRNA and some mitochondrial proteins

    • Jobs:

      • Energy metabolism (ATP synthesis)

        • Energy stored in the proton gradient drives ATP synthesis

      • Intracellular Ca2+ reservoir (important for cell signaling)

      • Important in apoptosis 

        • Cytochrome C is usually in the inner membrane but when released it starts apoptosis so no necrotising cell that burst this prevents inflammation.

  • Nucleus

    • Holds the genome and has the machinery necessary to maintain, copy and transcribe DNA.

    • Nuclear membrane is a double membrane the outer one is continuous with the  rough ER and the inner membrane is smooth and connected to the nuclear lamina (intermediate filiments)

      • Lamina are structural support and rebuild the nuclear envelope in divided cells

    • Nuclear pores (look like flowers)

      • Are used to let mRNA out and let hormones and proteins (need a specific sequence to get in, NLS Nuclear locator signal) in

  • Nuclear Chromatin

    • Complex of DNA and DNA binding proteins in the nucleus is chromatin

      • Euchromatin: actively coding spread out

        • Near nuclear interior

      • Heterochromatin: tightly packed NOT transcribing

        • Hetero men do nothing so Heterochromatin does nothing either and is not transcribed

        • Near the envelope and nucleolus 

    • Chromosomes are only visible as a distinct structure during Mitosis and Meiosis

    • During Interphase chromosomes occupy territories of the nucleus

  • Nucleolus 

    • Cells ribosomes production factory

      • Assemble ribosomes in there

    • Not really a body more of a designated area dedicated to code for rRNA

    • Proteins need to be brought in after ribosomes made in nucleus because NO proteins are synthesised in the nucleus only the ER and cytoplasm

  • Ribosomes:

    • Complex of two protein subunits and ribosomal RNA

    • Free floating or bought to ER

  • Endoplasmic Reticulum (ER)

    • Biggest organelles in the cell

    • Cells factories for secreted proteins and lipids

    • One network of membrane enclosed tubules and sacs (Cisternae) continuous membrane

      • 10% of total cell volume

    • Extends from the nuclear envelope throughout cytoplasm 

    • The orientation of transmembrane proteins is established in the ER and maintained as they travel along the secretory pathway.

      • Transmembrane proteins CANNOT flip sides

    • Proteins and lipids are packaged in vesicles and transported from the ER to the Golgi 

    • Smooth:

      • Contains enzymes for lipid, steroid and glycogen metabolism 

      • Detoxification 

        • The liver has a lot of smooth ER

      • Calcium Reservoir 

        • Important for muscle contractions

    • Rough:

      • Ribosomes

      • Role in synthesis of secreted and membrane proteins and protein folding and posttranslational modification

      • Folding of proteins in the ER with the help of Chaperones

      • Chaperones: ATP-Hydrolyzing enzyme refolds heat damaged proteins. Folds proteins into a tertiary structure which is not energetically favorable but biologically favorable    

      • Initial glycosylation of proteins happens in the ER (important for finishing of posttranslational modification in the golgi apparatus)

        • Manose is important in post translational modification

      • Add GPI anchors here for anchoring proteins of the cell membrane

  • Free floating ribosome proteins go to

    • Nucleus

    • Mitochondria

    • Peroxisomes

  • Bond ribosomal proteins go to:

    • Nuclear membrane

    • Golgi

    • Plasma membrane

    • Secretory vesicle 

    • Endosomes

    • Lysosomes

    • Peroxisome membrane

  • Golgi Apparatus

    • The “finishing shop” for proteins and lipids

    • Made up of: cis-Golgi (close to the RER), medial-Golgi, trans-Golgi, and trans-Golgi network (the end facing the plasma membrane)

    • Job

      • Protien maturation

      • TARGET PROTEINS TO THEIR APPROPRIATE SUBCELLULAR LOCATION.

      • Recycling proteins

      • Synthesis of glycolipids and sphingomyelin (important in myelin sheath of neurons)

    • Type of modifications done in the Golgi determine the final destination after release from the golgi

    • Proteins destined for LYSOSOMES are modified by MANNOSE phosphorylation

    • In polarized cells proteins are sorted into distinct transport vesicles to reach specific plasma membrane domains

  • Lysosomes

    • Digestive organelles of a cell

    • Acidic so low pH

    • Acid hydrolases inside to help breakdown cellular debris and ingest materials

    • Proton pump makes the interior extremely acidic to help protein breakdown

    • Autophagy: self eating (done to old or damaged cell structures) MAINTAINS CELLULAR HOMEOSTASIS

    • Heterophage: degrading foreign material

    • LYSOSOME FORMS FROM THE TRANS-GOLGI NETWORK

    • ENDOSOMES BECOMES PHAGOSOMES

  • Peroxisomes

    • Peroxisomes because they have a lot of Peroxide in it

    • Oxidases: produces hydrogen peroxide

    • Catalase: neutralizes the hydrogen peroxide

    • Job

      • Lipid Metabolism 

        • Myelin sheath of nerve cells

      • Oxidative reactions and detoxification

        • Breaks down various substrates

      • Metabolism of reactive oxygen species (ROS)

        • Reducing oxidative stress

  • Proteasome

    • Think of it as a paper shredder

    • ATP dependant

    • Recycles proteins marked with POLYUBIQUITIN CHAINS

      • Regulatory proteins that need to be inactive

      • Misfolded proteins 

      • Non functional proteins 

      • Viral proteins

  • Endocytosis: Comes into the cell

  • Extocytosis: Leaves the cell

  • Pinocytosis: drinking

  • Transcytosis: goes completely through the cell from one side to the other

  • Phagocytosis: eating

  • Extracellular Matrix

    • Fills space between cells and connects tissues together

    • Produced by cells in the ECM

    • Composed of secreted proteins and polysaccharides but varies by tissue type

    • Job

      • Structural and mechanical support

      • Pathways for cell regulation

      • Important in embryonic development

      • Provides survival signals and sequesters growth factors

        • Vital for tissue pathology and repair

    • Key components:

      • Matrix structural proteins (Collagen and Elastin) MACROMOLECULES

      • Matrix Polysaccharides (Glucosaminoglycans (GAGs), protoglycans) MACROMOLECULES

        • GAGs

          • Ground substance surround collagen and glycoproteins 

          • Resists compressive forces on the matrix 

          • Permits rapid diffusion of nutrients between blood and tissue cells

          • Bond ALOT of water (jello in Joints)

          • Mediates growth factor signaling

            • Also modulates cell signaling in general

          • Example: Hyaluronic Acid (Hyaluronan) LEGEND HAT which are used for intrajoint injections to help joint issues by increasing ground substance; it can be used on all animal types because no protein components so no immune response.

          • Proteoglycans

            • Bottle brush structure

            • Protein core (in cartilage)

            • HUGE molecules

            • Form aggregates with other GAGs matrix proteins 

            • Core protein 5% mass and Pollysaccharide chain 95% mass

      • Adhesion proteins/glycoproteins (Fibronectin, laminin)

      • Cell Matrix interaction proteins (Integrins)

  • Collagen

    • MAJOR PROTEIN IN ECM

    • Resist tensile forces

    • 3  alpha chains braided together so depending on the strands braided there is a lot of variability

    • RICH in glycine, proline, and hydroxyproline 

    • Provides structural framework for ECM

    • Proline and lysine need to be hydroxylated in ER  before use because it ensures proper folding of the helices

      • Vitamin C dependant process (scurvy)

    • Lysyl-oxidase enzyme crosslinks hydroxylysine for tensile strength

    • Without crosslinks you don't have a strong fiber 

    • Collagen types:

      • Fibrillar

        • High tensile strength 

        • Plywood like arrangement of the fibers

        • Found in loose connective tissues, tendons, bones, and cornea

      • Sheet-forming

        • Form nets that surround organs

        • Basal lamina beneath epithelia and muscles and nerve cells made of TYPE 4 COLLAGENS

        • Descemet membrane found in the cornea and needs to be see through so hexagonal pattern

      • Anchoring/Linking

        • Connect linear collagen fibers to other ECM components 

        • Basement membrane

  • Elastin

    • Second most important after collagen 

    • Tissue elasticity 

    • 5x more extensible then a rubber band

    • 50% of the dry weight of the Aorta 

    • Rich in proline so it can twist up

    • Fibrillin microfibrils scaffold with deposited crosslinked elastin

  • Adhesion Proteins

    • Link matrix components together and then to cells

    • Fibonectin and laminin make it up

    • Fibronectins are relatively small compared to collagen and elastin which are huge molecules

    • Fibronectin binds to integrins which are bound to the intracellular actin of the cytoskeleton so that's how they connect to the cells

    • Fibronectin is important in wound healing and cell migration

    • Laminin is essential for basal lamina

    • Integrins

      • TRANSMEMBRANE receptors connect to ECM with the cytoskeleton 

      • Integrins attach cells to the ECM and link them to the cytoskeleton 

      • Integrin function at:

        • Hemidesmosomes: provides stable adhesion and resistance to mechanical stress. Connected to intermediate filaments

        • Focal adhesion points: ECM dependant signaling to cells as well as cell migration, connect to Factin 

  • Basal lamina

    • Basement membrane

    • Structural support

    • Major components are laminin (primary organizer of the sheet structure, three polypeptide chains alpha beta and y in crosslinked structure) and type 4 collagen (flexible and give the basal lamina tensile strength)

    • Integrins attach to lamin which attach to type 4 collagen which make fish net pattern.

  • Resident cells:

    • Fibroblasts: make most collagen, elastic fibers, and proteoglycans

    • Chondrocytes: secretes cartilage matrix

    • Osteoblasts: Produce calcified bone matrix

    • White and Brown fat cells

  • Transient cells:

    • Cells that move through the cell matrix but do not originate there example white blood cells

  • Both transient and resident cells need to be able to degrade the ECM for growth and movement. Resident cells also make ECM.

  • Matrix metalloproteases (MMPs) degrades the ECM 

  • Tissue inhibitors of MMPs or TIMPs stop the MMPs (resident cells secrete these)

LEARNING OBJECTIVES:

1. List the principal membrane-bound organelles in an animal cell and state their primary functions.

Plasma membrane: To act as a selective permeable barrier that separates the cells internal environment from the outside. While regulating the movement of substance in and out.

Cytoskeleton: To provide structural support, shape, and internal organization while enabling cell movement and material transport.

Tight Junctions: To create a water tight semipermeable seal between adjacent cells that blocks movement of fluids, ions, and molecules through the space between them.

Anchoring junctions (Adherin junctions, desmosomes, and semidesmosomes): To securely connect neighboring cells to one another or to the extracellular matrix, providing mechanical strength and structural integrity to tissues.

Gap Junctions: To provide a direct channel for communication between neighboring cells by allowing the movement of ions, water, and small molecules.

Nucleus: Storage of genetic material DNA and managing all cellular activity

Nucleolus: Produce and assemble ribosomes

Ribosomes: Synthesize proteins by translating instructions from mRNA

RER: Protein synthesis , folding and modifications of proteins destined for cell membrane, secretion, or use within specialized organelles. 

SER: Synthesis of lipids and phospholipids and steroid hormones.

Golgi Apparatus: To modify and sort and package proteins and lipids received from the ER so they can be used inside the cell or sent outside of it.

Lysosomes: To act as the cell's garbage disposal and recycling center by breaking down waster old cell parts and foreign invaders.

Peroxisomes: Lipid metabolism and detoxification of harmful substances through oxidation reactions (peroxide)

Proteasome: to degrade unneeded, damaged, or misfolded proteins by breaking their peptide bonds

Mitochondria: Power house of the cell ATP synthesis as well as Ca2+ storage and helps in apoptosis. 

2. Explain how microtubules, microfilaments, and intermediate filaments differ in structure and how each supports cell shape, transport, and mechanical strength.


3. Describe and differentiate between tight junctions, adherens junctions, desmosomes, hemidesmosomes, and gap junctions in terms of molecular composition and physiological role.


4. Assess how combined defects in cytoskeletal networks and junctional complexes could impair epithelial integrity and contribute to clinical signs in animals.


5. Construct a labeled schematic of a eukaryotic cell showing organelles, cytoskeletal filaments, and junctional complexes, accurately depicting their spatial relationships and interactions.


6. List the structural components of a mitochondrion and name three of its key functions.


7. Explain how the chemiosmotic gradient across the inner mitochondrial membrane drives ATP synthesis.


8. Differentiate euchromatin from heterochromatin in terms of DNA packing, transcriptional activity, and nuclear architecture, and explain the role of lamins, nuclear pores and the nucleolus in genome regulation.


9. Describe the role of rER in translation and organelle targeting of proteins. Explain which proteins require translation in the rER, and describe ER dependent posttranslational modification.


10. Explain the special role of mannosidase in ER mediated protein synthesis.


11.Describe the structural organization of the Golgi apparatus and its role in protein and lipid processing, including glycosylation and vesicle trafficking.


12. Explain how proteins are targeted to lysosomes via mannose-6-phosphate tagging and the role of the trans-Golgi network in this process.


13. Differentiate between apical and basolateral protein targeting in polarized epithelial cells.


14. Summarize the functions of lysosomes in autophagy, heterophagy, and cellular homeostasis.


15. Identify the metabolic functions of peroxisomes, including lipid metabolism, ROS detoxification, and B-oxidation of very long chain fatty acids.


16. Discuss the role of proteasomes in ATP-dependent protein degradation, including the significance of polyubiquitin tagging and implications for cell cycle regulation and viral defense.


17. List major structural components of the extracellular matrix-including glycosaminoglycans, proteoglycans, collagen types, elastin, and adhesive glycoproteins-and describe their specific functions.


18.  Describe how varying extracellular matrix compositions of different tissues varies (bone, cartilage, skin, basal lamina, and blood vessels) contribute to mechanical properties and physiological roles.


19. Describe the structure and functional importance of the basal lamina, including its key molecular components (e.g., laminin, collagen IV, etc.) and describe its biological function role in cell polarity and filtration.


20. Describe the role of matrix metalloproteases (MMPs), serine proteases and tissue Inhibitors of metalloproteases (TIMPs) for ECM remodeling. Why is remodeling important, and how does dysregulated degradation contribute to pathological conditions such as cancer, inflammation, and fibrosis?


WOOCLAP QUESTIONS:
1. Are sodium levels higher inside the cell or outside?

  • Outside

2.  Tell me the size, what they're made of, and used for the different filaments.


Microtubules 

  • Largest filaments 

  • A- and b- tubulin

  • Movement (flagella, cilia)

Microfiliment 

  • Smallest filament

  • Actin and Myosin

  • Elastic support 

    • Important in muscle contraction

    • Pinch cytoplasm to make daughter cells

Intermediate Filament 

  • In between microfilament and microtubules in size

  • Many different bases: keratin, GFAP, and Desmins…

  • Used to stabilize the cell shape


GAGs/Proteoglycans: Ground substance for matrix 

Collagen: Structural framework of ECM

Elastin: Elasticity (especially in the lungs and blood vessels)

Adhesion Proteins: Adhesion and Communication

Type 4 collagen: important in the basal lamina