BIOCHEMISTRY are continuously synthesized and degraded –

Chapter 9: Enzymes: Regulation of Protein Turnover

 Constitutive Proteins – whose concentrations
Activities
remain essentially constant over time

BIOMEDICAL IMPORTANCE
CONTROL OF ENZYME SYNTHESIS
 Claude Bernard – conceptual basis for
 Inducers – promotes synthesis of certain
metabolic regulation
enzymes; substrates that stimulate the
 Walter Cannon – coined the term
transcription of the gene that encodes them
“Homeostasis”
 An excess of a metabolite may curtail synthesis
 Vibrio cholera – causes Cholera; disables
of its cognate enzyme via Repression
sensor response pathways in intestinal epithelial
 Both induction and repression involve cis
cells by ADP-ribosylating the GTP-binding
elements, specific DNA sequences and trans-
proteins that link cell surface receptors to
acting regulatory proteins
adenylyl cyclase
 Transcription factors – synthesize other
 Yersinia pestis – causes Plague; elaborates a
enzymes; is controlled by the interaction of
protein-tyrosine phosphatase that hydrolyzes
hormones and other extracellular signals with
phosphoryl groups on key cytoskeletal proteins
specific cell-surface receptors
 Epigenetic – DNA-independent information
systems
CONTROL OF ENZYME DEGRADATION
 Aaron Ciechanover
REGULATION OF METABOLITE FLOW
Avram Hershko
 Responses to Substrate changes is through
Irwin Rose – earned a Nobel prize for the
Passive means
Ubiquitin-proteasome Pathway
 Regulate Enzyme efficiency is in an Active
 26S Proteasome where degradation takes
means
place; a large macromolecular subunits
 Metabolite flow tends to be Unidirectional; so
arranged in a form of a hollow cylinder
as many reversible reactions occur this way
 Proteins are targeted to the interior of the
proteasome by Ubiquination – covalent
COMPARTMENTATION
attachment of one or more ubiquitin molecules
 Ensures metabolic efficiency and simplifies
 Ubiquitin – 8.5 kDa protein that is highly
regulation
conserved among eukaryotes
 Rate Limiting Reaction – decreasing the
 Ubiquination – catalyzed by a large family of
catalytic efficiency or the quantity of the
enzymes called E3 ligases, which attach
catalyst responsible for the “bottleneck;”
ubiquitin to the side-chain amino group lysyl
reduces metabolic flux through the entire
residues
pathway
 Conversely increasing in quantity or catalytic
UBIQUITIN-PROTEASOME PATHWAY
efficiency will enhance flux through the pathway
 Responsible for the regulated degradation of
 Acetyl-CoA Carboxylase – catalyzes the
selected cellular proteins and for removal of
synthesis of malonyl-CoA, the 1st committed
defective or aberrant proteins
reaction of fatty acid biosynthesis
 Key to the versatility and selectivity resides in
 Statin Drugs – synthesis of cholesterol by
the variety of intracellular E3 ligases and their
inhibiting HMG-CoA reductase, catalyst of the
ability to discriminate between the different
rate-limiting reaction of Cholesterogenesis
physical and conformational states of target
proteins
REGULATION OF ENZYME QUANTITY
 Catalytic capacity can be controlled by
REGULATION OF CATALYTIC ACTIVITY
changing the quantity of enzymes present;
 Allosteric Regulation – changes in intrinsic
altering its intrinsic catalytic efficiency or
catalytic efficiency effected by binding of
combination of both
dissociable ligands
 Schoenheimer – Proteins exist in a state of
 Covalent Modification- also involved in
“Dynamic Equilibrium” in the body where they
changes
ALLOSTERIC EFFECTORS
 Negative Allosteric Effector – its ability to bind
and inhibit an enzyme
 Allosteric Site – where a Negative Allosteric
effector binds; proposed by Jacques Monod
 Cooperative Feedback Inhibition – where the
effect of an excess of two or more end products
may be strictly additive, greater than their
individual effect
 Effectors are not isosteric since they are not
structurally the same with substrates but rather
Allosteric
 Allosteric enzymes – catalysis at the active site
may be modulated by the presence of effectors
at an allosteric site
ASPARTATE TRANSCARBAMOYLASE – catalyst for
1st reaction to pyrimidine biosynthesis
 Target of feedback regulation by 2 nucleotide
triphosphates, cytidine triphosphate and ADP
 CTP inhibits ATCase, whereas ATP activates it
 High levels of ATP overcome inhibition of CTP
enabling synthesis of pyrimidine to proceed
when purine levels are elevated
FEEDBACK REGULATION
 Feedback Regulation – phenomenologic term
devoid of mechanistic implications
 Feedback Inhibition – mechanism for
regulation of enzyme activity; may be
competitive, noncompetitive, partially
competitive or mixed.
SECOND MESSENGERS
 Specialized allosteric effectors that elicit
changes in the rate of enzyme-catalyzed
reaction within target cells through nerve
impulses and the binding of many hormones to
cell surface receptors
REGULATORY COVALENT MODIFICATIONS
 Partial Proteolysis & Phosphorylation –
frequently employed to regulate catalytic
activity of enzymes
HISTONE CODE
 Represents a classical example of Epigenetics,
hereditary transmission of information by means
other than the sequence of nucleotides that
comprise the genome
PROTEASES
 Secreted as catalytically inactive proenzymes
 Proproteins – inactive precursor proteins
- Includes Insulin, pepsin, trypsin,
chymotrypsin, several blood clotting factors
an collagen
 Proenzymes/Zymogens – enzymes formed by
proproteins
 Proteolytic activation of proproteins constitutes a
physiologically irreversible modification
because reunification of the 2 portions of a
protein produced by hydrolysis of a peptide bond
is entropically disfavored
 Proteases as catalytically inactive proenzyme;
protect tissue of origin from autodigestion,
such as can occur in pancreatitis
REVERSIBLE COVALENT MODIFICATION
 Phosphorylation-Dephosphorylation &
Acetylation-Deacetylation – most common
among the covalent modification
 Protein kinase- phosphorylate proteins by
catalyzing transfer of the terminal phosphoryl
group of ATP
 Protein Phosphatase – unmodified form of the
protein can be regenerate by hydrolytic removal
of phosphoryl groups
PROTEIN ACETYLATION
 Associated with histones and other nuclear
proteins
 Lysine acetyltransferase – catalyze the
transfer of the acetyl group of acetyl-CoA to the
E-amino groups of lysil residues, forming N-
acetyl lysine
2 Classes of Protein Deacetylases
o Histone Deacetylase – removal by
hydrolysis of acetyl groups
o Sirtuins – use NAD+ as substrate
PROTEIN PHOSPHORYLATION
 Highly versatile and selective process
 Many protein kinase and most protein
phosphatases act on more than one protein and
are interconverted between active and inactive
forms by the binding of second messengers or
by covalent modification by Phosphorylation
 Provide the basis for regulatory networks
that integrate multiple environmental input
signals to evoke appropriate coordinated
cellular response
CHECKPOINTS
 Elaborate monitoring systems asses as key
indicators of progress to ensure that no phase of
the cycle is initiated until the prior phase is
complete