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🧬Proteomics

Significant Protein Degradation Pathways

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Why This Matters

Protein degradation isn't just cellular housekeeping—it's a tightly regulated system that controls nearly every aspect of cell biology. When you're tested on proteomics, you need to understand that cells don't simply make proteins and let them accumulate forever. Instead, selective protein turnover drives the cell cycle, triggers apoptosis, responds to stress, and maintains the delicate balance we call proteostasis. Diseases from cancer to neurodegeneration often stem from degradation pathways gone wrong, making this content highly testable.

Don't just memorize pathway names—know why each pathway exists and what cellular problem it solves. Ask yourself: Is this pathway targeting damaged proteins? Regulating signaling? Responding to stress? Understanding the underlying logic will help you tackle comparison questions and FRQs that ask you to explain how cells maintain protein homeostasis under different conditions.


Ubiquitin-Dependent Degradation Systems

The ubiquitin system is the cell's primary "tag and destroy" mechanism. Ubiquitin, a small 76-amino acid protein, gets covalently attached to target proteins through an enzymatic cascade (E1, E2, E3 enzymes), marking them for destruction by the proteasome.

Ubiquitin-Proteasome System (UPS)

  • Polyubiquitin chains—specifically K48-linked chains—serve as the canonical "eat me" signal that directs proteins to the 26S proteasome
  • The 26S proteasome is a barrel-shaped complex with a 20S catalytic core flanked by 19S regulatory caps that unfold and thread substrates through for degradation
  • Regulates cell cycle progression by degrading cyclins and CDK inhibitors at precise times, making UPS dysfunction a hallmark of many cancers

N-end Rule Pathway

  • N-terminal amino acid identity determines protein half-life—destabilizing residues (Arg, Lys, Phe) trigger rapid ubiquitination while stabilizing residues (Met, Ser) promote longevity
  • N-recognins are the E3 ubiquitin ligases that recognize these destabilizing N-terminal residues and initiate degradation
  • Regulates protein turnover in response to cellular conditions, including oxygen sensing and cardiovascular development

Sumoylation-Dependent Protein Degradation

  • SUMO (Small Ubiquitin-like Modifier) attachment can either protect proteins from degradation or, paradoxically, target them for destruction via SUMO-targeted ubiquitin ligases (STUbLs)
  • Cross-talk with ubiquitination creates complex regulatory networks where SUMOylation status influences whether a protein gets degraded
  • Critical for stress responses and DNA damage repair, where SUMOylation helps clear damaged proteins from chromatin

Compare: UPS vs. N-end Rule Pathway—both rely on ubiquitination and proteasomal degradation, but UPS responds to diverse signals (phosphorylation, oxidation) while the N-end rule specifically reads the N-terminal amino acid as a degradation code. If asked about how cells regulate protein stability based on intrinsic protein features, the N-end rule is your example.


Lysosomal Degradation Pathways

When cells need to degrade large structures—entire organelles, protein aggregates, or bulk cytoplasm—they turn to the lysosome. These acidic compartments (pH ~4.5-5) contain hydrolases that break down proteins, lipids, and carbohydrates delivered through various autophagy mechanisms.

Autophagy-Lysosomal Pathway (Macroautophagy)

  • Double-membrane autophagosomes engulf cytoplasmic cargo (damaged organelles, protein aggregates) and fuse with lysosomes for degradation
  • LC3 lipidation (LC3-I to LC3-II conversion) is the key biochemical marker used to monitor autophagy activation in experiments
  • Responds to nutrient deprivation by recycling cellular components for energy and building blocks, making it essential for survival during starvation

Chaperone-Mediated Autophagy (CMA)

  • KFERQ-like motifs in target proteins are recognized by the cytosolic chaperone Hsc70, which delivers substrates directly to lysosomes
  • LAMP-2A receptor on the lysosomal membrane unfolds and translocates individual proteins into the lysosome—no autophagosome formation required
  • Selective degradation allows cells to remove specific damaged proteins without the bulk destruction of macroautophagy

Compare: Macroautophagy vs. CMA—both deliver cargo to lysosomes, but macroautophagy engulfs bulk cytoplasm non-selectively (or selectively via adaptor proteins), while CMA targets individual proteins containing specific sequence motifs. For FRQs about selective vs. bulk degradation, this distinction is critical.


Quality Control Pathways

Cells constantly monitor protein folding status and compartment integrity. These surveillance systems ensure that misfolded or damaged proteins are recognized and eliminated before they can aggregate or cause toxicity.

Endoplasmic Reticulum-Associated Degradation (ERAD)

  • Retrotranslocation moves misfolded ER proteins back through the Sec61 translocon into the cytosol, where they're ubiquitinated and degraded by the proteasome
  • ER quality control lectins (calnexin, calreticulin) and the EDEM proteins identify terminally misfolded glycoproteins for ERAD targeting
  • Prevents ER stress by clearing proteins that fail to fold properly; ERAD dysfunction triggers the unfolded protein response (UPR) and can lead to cell death

Heat Shock Protein-Mediated Degradation

  • Hsp70 and Hsp90 chaperones make triage decisions—they either refold damaged proteins or, if refolding fails, direct them to the proteasome or autophagy
  • CHIP (C-terminus of Hsp70-Interacting Protein) is an E3 ligase that ubiquitinates chaperone-bound clients destined for degradation
  • Activated during cellular stress (heat, oxidative damage) to prevent toxic protein aggregation—a key defense mechanism in neurodegenerative disease

Mitochondrial Protein Degradation

  • Compartmentalized proteases (Lon, ClpXP in the matrix; i-AAA and m-AAA in the inner membrane) degrade damaged mitochondrial proteins locally
  • Mitophagy eliminates entire damaged mitochondria via PINK1/Parkin-mediated ubiquitination of outer membrane proteins
  • Prevents oxidative stress by removing dysfunctional respiratory chain components that would otherwise generate reactive oxygen species

Compare: ERAD vs. Mitochondrial Degradation—both are organelle-specific quality control systems, but ERAD exports substrates to the cytosolic proteasome while mitochondria maintain their own internal proteases. This compartmentalization reflects the bacterial ancestry of mitochondria.


Programmed Proteolysis in Cell Fate

Some degradation pathways don't just maintain homeostasis—they execute irreversible cellular decisions. These proteases cleave specific substrates to trigger signaling cascades, cell death, or tissue remodeling.

Caspase-Mediated Proteolysis

  • Cysteine proteases cleaving after aspartate residues—this specificity (Asp in P1 position) allows caspases to target a defined set of ~1,000 substrates during apoptosis
  • Initiator caspases (8, 9, 10) activate executioner caspases (3, 6, 7) in a proteolytic cascade that dismantles the cell systematically
  • Cleaves structural proteins and DNases to produce the hallmarks of apoptosis: membrane blebbing, chromatin condensation, and DNA fragmentation

Calpain-Mediated Proteolysis

  • Calcium-dependent activation means calpains respond to signaling events that raise intracellular Ca2+Ca^{2+} concentrations
  • Limited proteolysis rather than complete degradation—calpains make specific cuts that modify protein function or generate active fragments
  • Critical for muscle remodeling and synaptic plasticity; dysregulated calpain activity contributes to muscular dystrophy and neurodegeneration

Compare: Caspases vs. Calpains—both are cysteine proteases that perform limited proteolysis, but caspases are activated by apoptotic signals and commit cells to death, while calpains respond to calcium signals and typically modulate protein function without killing the cell. Know which pathway to cite when asked about apoptosis (caspases) vs. signal-dependent protein modification (calpains).


Quick Reference Table

ConceptBest Examples
Ubiquitin-dependent proteasomal degradationUPS, N-end Rule, ERAD
Lysosomal degradationMacroautophagy, CMA
Organelle quality controlERAD (ER), Mitochondrial proteases, Mitophagy
Chaperone-assisted degradationHsp70/CHIP pathway, CMA
Apoptotic proteolysisCaspases
Signal-dependent proteolysisCalpains (calcium), Caspases (death signals)
Stress-responsive pathwaysAutophagy, HSP-mediated, UPS
Ubiquitin-like modifier systemsSUMOylation-dependent degradation

Self-Check Questions

  1. Which two degradation pathways both deliver substrates to lysosomes, and what distinguishes the selectivity of each mechanism?

  2. A cell accumulates misfolded proteins in the ER lumen. Which pathway handles this, and what happens if this pathway fails?

  3. Compare caspase-mediated and calpain-mediated proteolysis: what activates each, and what cellular outcomes do they typically produce?

  4. You observe increased LC3-II levels and decreased LAMP-2A expression in stressed cells. What does this suggest about which autophagy pathway(s) are active?

  5. An FRQ asks you to explain how cells maintain proteostasis during heat stress. Which pathways would you discuss, and what role does each play in preventing protein aggregation?