David L. Nelson and Michael M.

Cox

LEHNINGER
PRINCIPLES OF BIOCHEMISTRY
Fifth Edition

CHAPTER 6

Enzymes

2008 W. H. Freeman and Company

Alivingsystemcontrolsitsactivitythroughenzymes
Anenzymeisaproteinmoleculethatisabiologicalcatalyst
withthreecharacteristics:
thebasicfunctionofanenzymeistoincreasetherateof
areaction
mostenzymesactspecificallywithonlyonereactant
(calledasubstrate)toproduceproducts.
enzymesareregulatedfromastateoflowactivitytohigh
activityandviceversa
Theindividualityofalivingcellisdueinlargeparttothe
uniquesetofsome3,000enzymesthatitisgenetically
programmedtoproduce.Ifevenoneenzymeismissingor
defective,theresultscanbedisastrous.

Theactivityofanenzymedepends,attheminimum,onaspecific
proteinchain.
Inmanycases,theenzymeconsistsoftheproteinandacombinationof
oneormorepartscalledcofactors.
Apoenzyme:Thepolypeptideorproteinpartoftheenzymeiscalled
andmaybeinactiveinitsoriginalsynthesizedstructure(Theinactive
formoftheapoenzymeisknownasaproenzymeorzymogen)
Anapoenzymetogetherwithitscofactor(s)iscalledaholoenzyme

substrate
Bindingsite

Reaction coordinate diagram.

Complementary shapes of a substrate and its binding site

on an enzyme.

Lysozyme reaction

Gambaranpemutusanikatanolehenzymedanenergiyangmenyertai

DerivationofMichaelisMentenEquation

WecannowexpressV0intermsof[ES].
SubstitutingtherightsideofEquation619for
[ES]inEquation611gives

A double-reciprocal of Michaelis Menten

equation or Lineweaver-Burk plot

Enzymeinhibitions
Reversible
Competitive(usedfordrugbinding/medicationtechnique)
Uncompetitive
Mixed
Irreversible(usedfordrugdesign)

Ex.Ethanoldehydrogenase

Irreversible inhibition

FactoraffectingEnzymes

substrate concentration
pH
temperature
inhibitors

The effect of change in concentration

(think about Michaelis-Menten Curve)
Enzyme concentration: at low enzyme concentration there is
great competition for the active sites and the rate of
reaction is low. As the enzyme concentration increases,
there are more active sites and the reaction can proceed at
a faster rate.
Eventually, increasing the enzyme concentration beyond a
certain point has no effect because the substrate
concentration becomes the limiting factor.
Substrate concentration: at a low substrate concentration
there are many active sites that are not occupied. This
means that the reaction rate is low.
When more substrate molecules are added, more enzymesubstrate complexes can be formed. As there are more
active sites, and the rate of reaction increases.
Eventually, increasing the substrate concentration yet
further will have no effect. The active sites will be
saturated so no more enzyme-substrate complexes can be
formed.

Properties of Enzymes relating to

their tertiary structure.

The activity of enzymes is strongly affected by

changes in pH and temperature. Each enzyme
works best at a certain pH and temperature,its
activity decreasing at values above and below
that point. This is because of the importance of
tertiary structure (i.e. shape) in enzyme function
and forces, e.g., ionic interactions and hydrogen
bonds, in determining that shape.

ThepHdependencyofenzyme

The effect of pH
Extreme pH levels will produce denaturation
The structure of the enzyme is changed
The active site is distorted and the substrate
molecules will no longer fit in it
At pH values slightly different from the enzymes
optimum value, small changes in the charges of
the enzyme and its substrate molecules will
occur
This change in ionisation will affect the binding of
the substrate with the active site.

The effects of change in

temperature.
Temperature: enzymes work best at an optimum
temperature.
Below this, an increase in temperature provides more
kinetic energy to the molecules involved. The numbers of
collisions between enzyme and substrate will increase so
the rate will too.
Above the optimum temperature, and the enzymes are
denatured. Bonds holding the structure together will be
broken and the active site loses its shape and will no
longer work

The effect of Inhibitors

Lookbacktoenzymeinhibitions!!!

Allostericenzymes
Allostericenzymesareenzymesthatchangetheir
conformationalensembleuponbindingofaneffector,which
resultsinanapparentchangeinbindingaffinityatadifferent
ligandbindingsite.
This"actionatadistance"throughbindingofoneligand
affectingthebindingofanotheratadistinctlydifferentsite,is
theessenceoftheallostericconcept.
Allosteryplaysacrucialroleinmanyfundamentalbiological
processes,includingbutnotlimitedtocellsignalingandthe
regulationofmetabolism

Whereas enzymes without coupled domains/subunits display

normal MichaelisMenten kinetics, most allosteric enzymes
havemultiplecoupleddomains/subunitsandshowcooperative
binding. Generally speaking, such cooperativity results in
allosteric enzymes displaying a sigmoidal dependence on the
concentration of their substrates in positively cooperative
system
Thisallowsmostallosteric
enzymestogreatlyvarycatalytic
outputinresponsetosmall
changesineffectorconcentration.

Subunit interactions in an allosteric enzyme, and

interactions with inhibitors and activators.

Feedbackinhibition

(importantforcontrollingmetabolims)

Theinhibitionitselfcanbereversible
inhibitionsorthroughallosteric
modulation

Regulation of muscle glycogen phosphorylase

activity by multiple mechanisms

Itinvolvesallostericregulation
Andcascademechanismregulatedbyhormone

phosphoprotein phosphatase 1 (PP1)

phosphoprotein phosphatase inhibitor 1 (PPI-1)
phosphoprotein phosphatase 2B (PP2B)
cAMP-dependent protein kinase (PKA).

EndofEnzymeslecture

Enzymes are the tools of life