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Chapter 8
CYTOPLASMIC MEMBRANE
SYSTEMS: STRUCTURE,
FUNCTION, AND MEMBRANE
TRAFFICKING
Endomembrane System
 Endoplasmic reticulum
 Golgi complex
 Endosomes
 Lysosomes
 Vacuoles
8.1 An Overview of the
     Endomembrane System
 The organelles of the endomembrane system
 are part of a dynamic, integrated network in
 which materials are shuttled back and forth
 from one part of the cell to another.
Pathways:
 Biosynthetic (or Secretory) Pathway
  Moves proteins from their site of
   synthesis in the ER through the Golgi
   complex to the final destination

 Endocytic Pathway
  Moves material in the opposite direction
   from the plasma membrane or
   extracellular space to the cell interior.
Types of Secretory
Activities of Cells:
 Constitutive Secretion
  Materials are transported in secretory
   vesicles from their sites of synthesis and
   discharged into the extracellular space in a
   continual matter.

 Regulated Secretion
  Materials are stored as membrane-bound
   packages and discharged only in response to
   an appropriate stimulus.
Cytoplasmic membrane systems (chap8)
Cytoplasmic membrane systems (chap8)
8.2 A FEW APPROACHES TO
       THE STUDY OF
      ENDOMEMBRANES

 Determining the functions of
 cytoplasmic organelles required
 the development of new
 techniques and the execution of
 innovative experiments
Techniques:
 Autoradiography
 Green Fluorescent Protein (GFP)
 Biochemical Analysis of
  Subcellular Fractions
 Cell-Free Systems
 Study of Mutant Phenotypes
Autoradiography
 Utilized by James Jamieson and George
 Palade of Rockefeller University

 Provides a means to visualize
 biochemical processes by allowing an
 investigator to determine the location
 of radioactively labeled materials
 within a cell
 Tissue sections containing radioactive
 isotopes are covered with a thin layer
 of photographic emulsion, which is
 exposed by radiation emanating from
 radioisotopes within the tissue

 Sites in the cells containing
 radioactivity are revealed under the
 microscope by silver grains in the
 overlying emulsion
Using this
approach, the
endoplasmic
reticulum was
discovered to
be the site of
synthesis of
secretory
proteins
“pulse-chase”

 Pulse – refers to the brief incubation with
  radioactivity during which labeled amino
  acids are incorporated into protein

 Chase – refers to the period when the
  tissue is exposed to the unlabeled
  medium, a period during which additional
  proteins are synthesized using
  nonradioactive amino acids
 The longer the chase, the farther the
 radioactive proteins manufactured
 during the pulse will have traveled
 from their site of synthesis within the
 cell
Green Fluorescent Protein
          (GFP)
 Utilizes a gene isolated from a
  jellyfish that encodes a small protein
  which emits a green fluorescent light

 The use of GFP reveals the
 movement of proteins within a living
 cell
Cytoplasmic membrane systems (chap8)
Biochemical Analysis of
 Subcellular Fractions

 Techniques to break up
 (homogenize) cells and isolate
 particular types of organelles
 were pioneered by Albert Claude
 and Christian De Duve
Cell-Free Systems
 Did not contain whole cells

 Provided a wealth of
 information about complex
 processes that were impossible
 to study using intact cells
 Researchers have used
 cell-free systems to
 identify the roles of
 many of the proteins
 involved in membrane
 trafficking
Study of Mutant Phenotypes

 Mutant = organism (or cultured cell)
 whose chromosomes contain one or
 more genes that encode abnormal
 proteins

 When a protein encoded by a mutant
 gene is unable to carry out its normal
 function, the cell carrying the mutation
 exhibits a characteristic deficiency
8.3 The Endoplasmic
     Reticulum (ER)

 A system of tubules, cisternae,
 and vesicles that divides the
 cytoplasm into a luminal (or
 cisternal) space within the ER
 membranes and a cytosolic
 space outside the membranes.
ER Subcompartmets:
 Rough Endoplasmic Reticulum (RER)
  Occurs typically as flattened cisternae and
   whose membranes have attached ribosomes

 Smooth Endoplasmic Reticulum (SER)
  Occurs typically as tubular compartments and
   whose membranes lack ribosomes
Smooth ER Functions:
 Synthesis of steroid hormones in the
  endocrine cells of the gonad and adrenal
  cortex

 Detoxification in the liver of a wide variety of
  organic compounds

 Sequestering calcium ions within the
  cytoplasm of cells
Rough ER Functions:
 Synthesis of secreted proteins, lysosomal
  proteins, and integral membrane proteins

 The RER is the starting point of the
  biosyntetic pathway: it is the site of
  synthesis of the proteins, carbohydrate
  chains, and phospholipids that journey
  through the membranous compartments
  of the cell.
 Proteins to be synthesized on
 membrane-bound ribosomes of the
 RER are recognized by a hydrophobic
 signal sequence, which is usually
 situated near the N-terminus of the
 nascent polypeptide.
Cytoplasmic membrane systems (chap8)
Glycosylation in the RER
 The addition of sugars (glycosylation)
 to the asparagine residues of proteins
 begins in the RER and continues in the
 Golgi complex.

 The addition of sugars to an
 oligosaccharide chain is catalyzed by a
 family of membrane-bound enzymes
 called glycosyltransferases.
8.4 The Golgi Complex
 Consist primarily of flattened, dislike,
  membranous cisternae with dilated rims and
  associated vesicles and tubules.

 Functions as a processing plant, modifying
  the membrane components and cargo
  synthesized in the ER before it moves on to
  its target destination.
 Divided into several
 functionally distinct
 compartments arranged along
 an axis from the cis or entry
 face closest to the ER to the
 trans or exit face at the
 opposite end of the stack.
Cytoplasmic membrane systems (chap8)
 The cis-most face of the organelle
 is composed of an interconnected
 network of tubules referred to as
 the cis Golgi network (CGN)

  Function primarily as a sorting station
   that distinguishes between proteins
   to be shipped back to the ER and
   those that are allowed to proceed to
   the next Golgi station.
 The trans-most face of the
 organelle contains a distinct
 network of tubules and vesicles
 called the trans Golgi network
 (TGN).

  The TGN is a sorting station where
   proteins are segregated into different
   types of vesicles heading either to the
   plasma membrane or to various
   intracellular destinations.
Cytoplasmic membrane systems (chap8)
 As the materials traverse the
 stack, sugars are added to
 the oligosaccharide chains
 by glycosyltransferases
 located in particular Golgi
 cisternae.
 The bulk of the Golgi complex consists
 of a series of large, flattened
 cisternae, which are divided into cis,
 medial, and trans cisternae.
 Once the proteins reach the
 trans Golgi network (TGN) at
 the end of the stack, they are
 ready to be sorted and
 targeted for delivery to their
 ultimate cellular or
 extracellular destination.

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Cytoplasmic membrane systems (chap8)

  • 1. Chapter 8 CYTOPLASMIC MEMBRANE SYSTEMS: STRUCTURE, FUNCTION, AND MEMBRANE TRAFFICKING
  • 2. Endomembrane System  Endoplasmic reticulum  Golgi complex  Endosomes  Lysosomes  Vacuoles
  • 3. 8.1 An Overview of the Endomembrane System  The organelles of the endomembrane system are part of a dynamic, integrated network in which materials are shuttled back and forth from one part of the cell to another.
  • 4. Pathways:  Biosynthetic (or Secretory) Pathway  Moves proteins from their site of synthesis in the ER through the Golgi complex to the final destination  Endocytic Pathway  Moves material in the opposite direction from the plasma membrane or extracellular space to the cell interior.
  • 5. Types of Secretory Activities of Cells:  Constitutive Secretion  Materials are transported in secretory vesicles from their sites of synthesis and discharged into the extracellular space in a continual matter.  Regulated Secretion  Materials are stored as membrane-bound packages and discharged only in response to an appropriate stimulus.
  • 8. 8.2 A FEW APPROACHES TO THE STUDY OF ENDOMEMBRANES  Determining the functions of cytoplasmic organelles required the development of new techniques and the execution of innovative experiments
  • 9. Techniques:  Autoradiography  Green Fluorescent Protein (GFP)  Biochemical Analysis of Subcellular Fractions  Cell-Free Systems  Study of Mutant Phenotypes
  • 10. Autoradiography  Utilized by James Jamieson and George Palade of Rockefeller University  Provides a means to visualize biochemical processes by allowing an investigator to determine the location of radioactively labeled materials within a cell
  • 11.  Tissue sections containing radioactive isotopes are covered with a thin layer of photographic emulsion, which is exposed by radiation emanating from radioisotopes within the tissue  Sites in the cells containing radioactivity are revealed under the microscope by silver grains in the overlying emulsion
  • 12. Using this approach, the endoplasmic reticulum was discovered to be the site of synthesis of secretory proteins
  • 13. “pulse-chase”  Pulse – refers to the brief incubation with radioactivity during which labeled amino acids are incorporated into protein  Chase – refers to the period when the tissue is exposed to the unlabeled medium, a period during which additional proteins are synthesized using nonradioactive amino acids
  • 14.  The longer the chase, the farther the radioactive proteins manufactured during the pulse will have traveled from their site of synthesis within the cell
  • 15. Green Fluorescent Protein (GFP)  Utilizes a gene isolated from a jellyfish that encodes a small protein which emits a green fluorescent light  The use of GFP reveals the movement of proteins within a living cell
  • 17. Biochemical Analysis of Subcellular Fractions  Techniques to break up (homogenize) cells and isolate particular types of organelles were pioneered by Albert Claude and Christian De Duve
  • 18. Cell-Free Systems  Did not contain whole cells  Provided a wealth of information about complex processes that were impossible to study using intact cells
  • 19.  Researchers have used cell-free systems to identify the roles of many of the proteins involved in membrane trafficking
  • 20. Study of Mutant Phenotypes  Mutant = organism (or cultured cell) whose chromosomes contain one or more genes that encode abnormal proteins  When a protein encoded by a mutant gene is unable to carry out its normal function, the cell carrying the mutation exhibits a characteristic deficiency
  • 21. 8.3 The Endoplasmic Reticulum (ER)  A system of tubules, cisternae, and vesicles that divides the cytoplasm into a luminal (or cisternal) space within the ER membranes and a cytosolic space outside the membranes.
  • 22. ER Subcompartmets:  Rough Endoplasmic Reticulum (RER)  Occurs typically as flattened cisternae and whose membranes have attached ribosomes  Smooth Endoplasmic Reticulum (SER)  Occurs typically as tubular compartments and whose membranes lack ribosomes
  • 23. Smooth ER Functions:  Synthesis of steroid hormones in the endocrine cells of the gonad and adrenal cortex  Detoxification in the liver of a wide variety of organic compounds  Sequestering calcium ions within the cytoplasm of cells
  • 24. Rough ER Functions:  Synthesis of secreted proteins, lysosomal proteins, and integral membrane proteins  The RER is the starting point of the biosyntetic pathway: it is the site of synthesis of the proteins, carbohydrate chains, and phospholipids that journey through the membranous compartments of the cell.
  • 25.  Proteins to be synthesized on membrane-bound ribosomes of the RER are recognized by a hydrophobic signal sequence, which is usually situated near the N-terminus of the nascent polypeptide.
  • 27. Glycosylation in the RER  The addition of sugars (glycosylation) to the asparagine residues of proteins begins in the RER and continues in the Golgi complex.  The addition of sugars to an oligosaccharide chain is catalyzed by a family of membrane-bound enzymes called glycosyltransferases.
  • 28. 8.4 The Golgi Complex  Consist primarily of flattened, dislike, membranous cisternae with dilated rims and associated vesicles and tubules.  Functions as a processing plant, modifying the membrane components and cargo synthesized in the ER before it moves on to its target destination.
  • 29.  Divided into several functionally distinct compartments arranged along an axis from the cis or entry face closest to the ER to the trans or exit face at the opposite end of the stack.
  • 31.  The cis-most face of the organelle is composed of an interconnected network of tubules referred to as the cis Golgi network (CGN)  Function primarily as a sorting station that distinguishes between proteins to be shipped back to the ER and those that are allowed to proceed to the next Golgi station.
  • 32.  The trans-most face of the organelle contains a distinct network of tubules and vesicles called the trans Golgi network (TGN).  The TGN is a sorting station where proteins are segregated into different types of vesicles heading either to the plasma membrane or to various intracellular destinations.
  • 34.  As the materials traverse the stack, sugars are added to the oligosaccharide chains by glycosyltransferases located in particular Golgi cisternae.
  • 35.  The bulk of the Golgi complex consists of a series of large, flattened cisternae, which are divided into cis, medial, and trans cisternae.
  • 36.  Once the proteins reach the trans Golgi network (TGN) at the end of the stack, they are ready to be sorted and targeted for delivery to their ultimate cellular or extracellular destination.