INHIBITOR ENZIM

Mauidzotussyarifah (105040213111059)

Dosen Pembimbing Dr.Ir.Moch.Dawam Maghfoer M.S

Kelas E Program Studi Agroekoteknologi Fakultas Pertanian Universitas Brawijaya

Abstrak

Secara umum suatu inhibitor adalah suatu zat kimia yang dapat menghambat atau memperlambat
suatu reaksi kimia. Banyak zat dapat mempengaruhi proses metabolisme dengan mempengaruhi
aktivitas enzim. Inhibitor enzim sangat penting di sini. Sebagian besar obat-obatan bertindak
sebagai inhibitor enzim. eksperimen Enzyme kinetic merupakan aspek pengembangan obat dan
pengujian prosedur. metabolit alam juga terlibat dalam proses regulasi sebagai inhibitor. Laju
reaksi enzim dapat diturunkan menggunakan berbagai jenis inhibitor enzim, yaitu: refersible
inhibitor dan irrefesible inhibitor.

1. PENDAHULUAN

Inhibitor merupakan zat atau senyawa yang menghambat fungsi enzim, inhibitor sering
digunakan sebagai obat. Contohnya adalah inhibitor yang digunakan sebagai obat aspirin. Aspirin
menginhibisi enzim COX-1 dan COX-2 yang memproduksi pembawa pesan peradangan
prostaglandin, sehingga ia dapat menekan peradangan dan rasa sakit. Namun, banyak pula inhibitor
enzim lainnya yang beracun. Sebagai contohnya, sianida yang merupakan inhibitor enzim ireversibel,
akan bergabung dengan tembaga dan besi pada tapak aktif enzim sitokrom c oksidase dan memblok
pernafasan sel.

2. Inhibitor Enzim
2.1 Macam – Macam Inhibistor
a. Irrefesible Inhibitor
Inhibitor irreversibel (tidak dapat kembali), yaitu terjadi setelah inhibitor mengikat
enzim, inhibitor yang tidak dapat dipisahkan dari sisi aktif enzim. Keadaan ini menyebabkan enzim
tidak dapat mengikat substrat atau inhibitor merusak beberapa komponen (gugus fungsi) pada sisi
katalitik molekul enzim. Inhibitor ireversibel biasanya memodifikasi kovalen enzim, dan karena itu
 hambatan tidak dapat dikembalikan. Inhibitor ireversibel sering mengandung kelompok fungsional
reaktif seperti mustard nitrogen, aldehida, haloalkanes, alkena, akseptor Michael, sulfonat fenil, atau
fluorophosphonates.
Penghambatan ireversibel berbeda dari inaktivasi enzim ireversibel. Inhibitor ireversibel
umumnya spesifik untuk satu kelas dari enzim dan tidak menonaktifkan semua protein, mereka tidak
berfungsi dengan menghancurkan struktur protein tetapi dengan secara khusus mengubah situs aktif
dari target mereka. Misalnya, ekstrim pH atau temperatur biasanya menyebabkan denaturasi dari
semua struktur protein, tapi ini merupakan efek non-spesifik.
b. Refesible Inhibitor
Reversibel inhiabitor mengikat enzim dengan interaksi non-kovalen seperti ikatan
hidrogen, interaksi hidrofobik dan ikatan ion. Beberapa obligasi yang lemah antara inhibitor dan situs
aktif bergabung untuk menghasilkan kuat dan spesifik mengikat. Berbeda dengan substrat dan
inhibitor ireversibel, inhibitor reversibel umumnya tidak mengalami reaksi kimia ketika terikat enzim
dan dapat dengan mudah dihilangkan dengan pengenceran atau dialysis. inhibisi kompetitif: substrat
(S) dan inhibitor (I) bersaing untuk situs aktif. Ada tiga macam enzim inhibitor reversible. Mereka
diklasifikasikan menurut pengaruh variasi konsentrasi substrat enzim di inhibitor, yaitu : competitive
inhibition, non competitive inhibition, dan uncompetitive inhibition.
c. Feedback inhibitor
Pada enzim, Inhibisi umpan balik atau lebih dikenal Feedback Inhibitor adalah penekanan
aktivitas enzim yang berpartisipasi dalam urutan reaksi oleh substansi dan disintesis oleh sebuah
produk dari urutan itu. Ketika produk terakumulasi dalam sel melampaui jumlah optimal,
produksinya menurun akibat penghambatan enzim yang terlibat dalam sintesisnya. Enzim tersebut
memiliki kemampuan untuk mengkatalisis reaksi dan bergantung pada substrat molekul selain
mereka (yang mana mereka bertindak untuk membentuk suatu produk), dikatakan berada di bawah
kendali alosterik.

3. KESIMPULAN
Secara umum suatu inhibitor adalah suatu zat kimia yang dapat menghambat atau
memperlambat suatu reaksi kimia. Banyak zat dapat mempengaruhi proses metabolisme dengan
mempengaruhi aktivitas enzim. Eksperimen Enzyme kinetic merupakan aspek pengembangan obat
dan pengujian prosedur. metabolit alam juga terlibat dalam proses regulasi sebagai inhibitor. Laju
reaksi enzim dapat diturunkan menggunakan berbagai jenis inhibitor enzim, yaitu: refersible inhibitor
dan irrefesible inhibitor.
DAFTAR PUSTAKA

Anonymous, 2011.http://en.wikipedia.org/wiki/Enzyme_inhibitor#Reversible_inhibitors.

Aonymous, 2011. http://id.wikipedia.org/wiki/Enzim#Inhibisi

Anonymous, 2011. http://www.britannica.com/EBchecked/topic/203717/feedback-inhibition

LAMPIRAN DAFTAR PUSTAKA

Enzyme inhibitor
An enzyme inhibitor is a molecule that binds to enzymes and decreases their activity. Since
blocking an enzyme's activity can kill a pathogen or correct a metabolic imbalance, many drugs
are enzyme inhibitors. They are also used as herbicides and pesticides. Not all molecules that
bind to enzymes are inhibitors; enzyme activators bind to enzymes and increase their enzymatic
activity.

The binding of an inhibitor can stop a substrate from entering the enzyme's active site and/or
hinder the enzyme from catalysing its reaction. Inhibitor binding is either reversible or
irreversible. Irreversible inhibitors usually react with the enzyme and change it chemically.
These inhibitors modify key amino acid residues needed for enzymatic activity. In contrast,
reversible inhibitors bind non-covalently and different types of inhibition are produced
depending on whether these inhibitors bind the enzyme, the enzyme-substrate complex, or both.
Many drug molecules are enzyme inhibitors, so their discovery and improvement is an active area of
research in biochemistry and pharmacology. A medicinal enzyme inhibitor is often judged by its
specificity (its lack of binding to other proteins) and its potency (its dissociation constant, which
indicates the concentration needed to inhibit the enzyme). A high specificity and potency ensure that a
drug will have few side effects and thus low toxicity.

Enzyme inhibitors also occur naturally and are involved in the regulation of metabolism. For example,
enzymes in a metabolic pathway can be inhibited by downstream products. This type of negative
feedback slows flux through a pathway when the products begin to build up and is an important way to
maintain homeostasis in a cell. Other cellular enzyme inhibitors are proteins that specifically bind to and
inhibit an enzyme target. This can help control enzymes that may be damaging to a cell, such as
proteases or nucleases; a well-characterised example is the ribonuclease inhibitor, which binds to
ribonucleases in one of the tightest known protein–protein interactions.[1] Natural enzyme inhibitors can
also be poisons and are used as defences against predators or as ways of killing prey.

Reversible inhibitors
Types of reversible inhibitors
Reversible inhibitors bind to enzymes with non-covalent interactions such as hydrogen bonds,
hydrophobic interactions and ionic bonds. Multiple weak bonds between the inhibitor and the active
site combine to produce strong and specific binding. In contrast to substrates and irreversible inhibitors,
reversible inhibitors generally do not undergo chemical reactions when bound to the enzyme and can be
easily removed by dilution or dialysis.

There are four kinds of reversible enzyme inhibitors. They are classified according to the effect
of varying the concentration of the enzyme's substrate on the inhibitor.[2]

 In competitive inhibition, the substrate and inhibitor cannot bind to the enzyme at the same
time, as shown in the figure on the left. This usually results from the inhibitor having an affinity
for the active site of an enzyme where the substrate also binds; the substrate and inhibitor
compete for access to the enzyme's active site. This type of inhibition can be overcome by
sufficiently high concentrations of substrate, i.e., by out-competing the inhibitor. Competitive
inhibitors are often similar in structure to the real substrate (see examples below).

 In uncompetitive inhibition, the inhibitor binds only to the substrate-enzyme complex, it should
not be confused with non-competitive inhibitors. Both Vmax and Km decrease (maximum
velocity decreases as a result of removing activated complex while binding efficiency increases
as a result of Le Chatelier's principle).
  In mixed inhibition, the inhibitor can bind to the enzyme at the same time as the enzyme's
substrate. However, the binding of the inhibitor affects the binding of the substrate, and vice
versa. This type of inhibition can be reduced, but not overcome by increasing concentrations of
substrate. Although it is possible for mixed-type inhibitors to bind in the active site, this type of
inhibition generally results from an allosteric effect where the inhibitor binds to a different site
on an enzyme. Inhibitor binding to this allosteric site changes the conformation (i.e., tertiary
structure or three-dimensional shape) of the enzyme so that the affinity of the substrate for the
active site is reduced.

 Non-competitive inhibition is a form of mixed inhibition where the binding of the inhibitor to
the enzyme reduces its activity but does not affect the binding of substrate. As a result, the
extent of inhibition depends only on the concentration of the inhibitor.

Types of irreversible inhibition

Irreversible inhibitors usually covalently modify an enzyme, and inhibition cannot therefore be
reversed. Irreversible inhibitors often contain reactive functional groups such as nitrogen
mustards, aldehydes, haloalkanes, alkenes, Michael acceptors, phenyl sulfonates, or
fluorophosphonates. These electrophilic groups react with amino acid side chains to form
covalent adducts. The residues modified are those with side chains containing nucleophiles such
as hydroxyl or sulfhydryl groups; these include the amino acids serine (as in DFP, right),
cysteine, threonine or tyrosine.[13]

Irreversible inhibition is different from irreversible enzyme inactivation. Irreversible inhibitors

are generally specific for one class of enzyme and do not inactivate all proteins; they do not
function by destroying protein structure but by specifically altering the active site of their target.
For example, extremes of pH or temperature usually cause denaturation of all protein structure,
but this is a non-specific effect. Similarly, some non-specific chemical treatments destroy protein
structure: for example, heating in concentrated hydrochloric acid will hydrolyse the peptide
bonds holding proteins together, releasing free amino acids.[14]

Irreversible inhibitors display time-dependent inhibition and their potency therefore cannot be
characterised by an IC50 value. This is because the amount of active enzyme at a given
concentration of irreversible inhibitor will be different depending on how long the inhibitor is
pre-incubated with the enzyme. Instead, kobs/[I] values are used,[15] wherekobs is the observed
pseudo-first order rate of inactivation (obtained by plotting the log of % activity vs. time) and [I]
is the concentration of inhibitor. The kobs/[I] parameter is valid as long as the inhibitor does not
saturate binding with the enzyme (in which case kobs = kinact).
Analysis of irreversible inhibition

Kinetic scheme for irreversible inhibitors

As shown in the figure to the left, irreversible inhibitors form a reversible non-covalent complex
with the enzyme (EI or ESI) and this then reacts to produce the covalently modified "dead-end
complex" EI*. The rate at which EI* is formed is called the inactivation rate or kinact. Since
formation of EI may compete with ES, binding of irreversible inhibitors can be prevented by
competition either with substrate or with a second, reversible inhibitor. This protection effect is
good evidence of a specific reaction of the irreversible inhibitor with the active site.

The binding and inactivation steps of this reaction are investigated by incubating the enzyme
with inhibitor and assaying the amount of activity remaining over time. The activity will be
decrease in a time-dependent manner, usually following exponential decay. Fitting these data to
a rate equation gives the rate of inactivation at this concentration of inhibitor. This is done at
several different concentrations of inhibitor. If a reversible EI complex is involved the
inactivation rate will be saturable and fitting this curve will give kinact and Ki.[16]

Another method that is widely used in these analyses is mass spectrometry. Here, accurate
measurement of the mass of the unmodified native enzyme and the inactivated enzyme gives the
increase in mass caused by reaction with the inhibitor and shows the stoichiometry of the
reaction.[17] This is usually done using a MALDI-TOF mass spectrometer. In a complementary
technique, peptide mass fingerprinting involves digestion of the native and modified protein with
a protease such as trypsin. This will produce a set of peptides that can be analysed using a mass
spectrometer. The peptide that changes in mass after reaction with the inhibitor will be the one
that contains the site of modification.
Special cases

Chemical mechanism for irreversible inhibition of ornithine decarboxylase by DFMO. Pyridoxal 5'-
phosphate (Py) and enzyme (E) are not shown. Adapted from [18]

Not all irreversible inhibitors form covalent adducts with their enzyme targets. Some reversible
inhibitors bind so tightly to their target enzyme that they are essentially irreversible. These tight-
binding inhibitors may show kinetics similar to covalent irreversible inhibitors. In these cases,
some of these inhibitors rapidly bind to the enzyme in a low-affinity EI complex and this then
undergoes a slower rearrangement to a very tightly bound EI* complex (see figure above). This
kinetic behaviour is called slow-binding.[19] This slow rearrangement after binding often involves
a conformational change as the enzyme "clamps down" around the inhibitor molecule. Examples
of slow-binding inhibitors include some important drugs, such methotrexate,[20] allopurinol,[21]
and the activated form of acyclovir.[22]

Enzim
Dari Wikipedia bahasa Indonesia

Enzim adalah biomolekul berupa protein yang berfungsi sebagai katalis (senyawa yang mempercepat
proses reaksi tanpa habis bereaksi) dalam suatu reaksi kimia organik.[1][2] Molekul awal yang disebut
substrat akan dipercepat perubahannya menjadi molekul lain yang disebut produk. Jenis produk yang
akan dihasilkan bergantung pada suatu kondisi/zat, yang disebut promoter. Semua proses biologis sel
memerlukan enzim agar dapat berlangsung dengan cukup cepat dalam suatu arah lintasan metabolisme
yang ditentukan oleh hormon sebagai promoter.

Enzim bekerja dengan cara bereaksi dengan molekul substrat untuk menghasilkan senyawa
intermediat melalui suatu reaksi kimia organik yang membutuhkan energi aktivasi lebih rendah,
sehingga percepatan reaksi kimia terjadi karena reaksi kimia dengan energi aktivasi lebih tinggi
membutuhkan waktu lebih lama. Sebagai contoh:

X + C → XC (1)

Y + XC → XYC (2)

XYC → CZ (3)

CZ → C + Z (4)

Meskipun senyawa katalis dapat berubah pada reaksi awal, pada reaksi akhir molekul katalis
akan kembali ke bentuk semula.

Sebagian besar enzim bekerja secara khas, yang artinya setiap jenis enzim hanya dapat bekerja
pada satu macam senyawa atau reaksi kimia. Hal ini disebabkan perbedaan struktur kimia tiap
enzim yang bersifat tetap. Sebagai contoh, enzim α-amilase hanya dapat digunakan pada proses
perombakan pati menjadi glukosa.

Kerja enzim dipengaruhi oleh beberapa faktor, terutama adalah substrat, suhu, keasaman,
kofaktor dan inhibitor. Tiap enzim memerlukan suhu dan pH (tingkat keasaman) optimum yang
berbeda-beda karena enzim adalah protein, yang dapat mengalami perubahan bentuk jika suhu
dan keasaman berubah. Di luar suhu atau pH yang sesuai, enzim tidak dapat bekerja secara
optimal atau strukturnya akan mengalami kerusakan. Hal ini akan menyebabkan enzim
kehilangan fungsinya sama sekali. Kerja enzim juga dipengaruhi oleh molekul lain. Inhibitor
adalah molekul yang menurunkan aktivitas enzim, sedangkan aktivator adalah yang
meningkatkan aktivitas enzim. Banyak obat dan racun adalah inihibitor enzim.

Laju reaksi enzim dapat diturunkan menggunakan berbagai jenis inhibitor enzim.

Inhibisi kompetitif

Pada inihibisi kompetitif, inhibitor dan substrat berkompetisi untuk berikatan dengan enzim.
Seringkali inhibitor kompetitif memiliki struktur yang sangat mirip dengan substrat asli enzim.
Sebagai contoh, metotreksat adalah inihibitor kompetitif untuk enzim dihidrofolat reduktase.
Kemiripan antara struktur asam folat dengan obat ini ditunjukkan oleh gambar di samping
bawah. Perhatikan bahwa pengikatan inhibitor tidaklah perlu terjadi pada tapak pengikatan
substrat apabila pengikatan inihibitor mengubah konformasi enzim, sehingga menghalangi
pengikatan substrat. Pada inhibisi kompetitif, kelajuan maksimal reaksi tidak berubah, namun
memerlukan konsentrasi substrat yang lebih tinggi untuk mencapai kelajuan maksimal tersebut,
sehingga meningkatkan Km.

Inhibisi tak kompetitif

Pada inhibisi tak kompetitif, inhibitor tidak dapat berikatan dengan enzim bebas, namun hanya
dapat dengan komples ES. Kompleks EIS yang terbentuk kemudian menjadi tidak aktif. Jenis
inhibisi ini sangat jarang, namun dapat terjadi pada enzim-enzim multimerik.

Inhibisi non-kompetitif

Inhibitor non-kompetitif dapat mengikat enzim pada saat yang sama substrat berikatan dengan
enzim. Baik kompleks EI dan EIS tidak aktif. Karena inhibitor tidak dapat dilawan dengan
peningkatan konsentrasi substrat, Vmax reaksi berubah. Namun, karena substrat masih dapat
mengikat enzim, Km tetaplah sama.

Inhibisi campuran

Inhibisis jenis ini mirip dengan inhibisi non-kompetitif, kecuali kompleks EIS memiliki aktivitas
enzimatik residual.

Pada banyak organisme, inhibitor dapat merupakan bagian dari mekanisme umpan balik. Jika
enzim memproduksi terlalu banyak produk, produk tersebut dapat berperan sebagai inhibitor
bagi enzim tersebut. Hal ini akan menyebabkan produksi produk melambat atau berhenti. Bentuk
umpan balik ini adalah umpan balik negatif. Enzim memiliki bentuk regulasi seperti ini sering
kali multimerik dan mempunyai tapak ikat alosterik. Kurva substrat/kelajuan enzim ini tidak
berbentuk hiperbola melainkan berbentuk S.

Koenzim asam folat (kiri) dan obat anti kanker metotreksat (kanan) memiliki struktur yang sangat mirip.
Oleh sebab itu, metotreksat adalah inhibitor kompetitif bagi enzim yang menggunukan folat.

Inhibitor ireversibel bereaksi dengan enzim dan membentuk aduk dengan protein. Inaktivasi ini
bersifat ireversible. Inhibitor seperti ini contohnya efloritina, obat yang digunakan untuk
mengobati penyakit yang disebabkan oleh protozoa African trypanosomiasis.[57] Penisilin dan
Aspirin juga bekerja dengan cara yang sama. Senyawa obat ini terikat pada tapak aktif, dan
enzim kemudian mengubah inhibitor menjadi bentuk aktif yang bereaksi secara ireversibel
dengan satu atau lebih residu asam amino.

Kegunaan inhibitor

Oleh karena inhibitor menghambat fungsi enzim, inhibitor sering digunakan sebagai obat.
Contohnya adalah inhibitor yang digunakan sebagai obat aspirin. Aspirin menginhibisi enzim
COX-1 dan COX-2 yang memproduksi pembawa pesan peradangan prostaglandin, sehingga ia
dapat menekan peradangan dan rasa sakit. Namun, banyak pula inhibitor enzim lainnya yang
beracun. Sebagai contohnya, sianida yang merupakan inhibitor enzim ireversibel, akan
bergabung dengan tembaga dan besi pada tapak aktif enzim sitokrom c oksidase dan memblok
pernafasan sel.[58]
feedback inhibition,  in enzymology, suppression of the activity of an enzyme, participating in a
sequence of reactions by which a substance is synthesized, by a product of that sequence. When
the product accumulates in a cell beyond an optimal amount, its production is decreased by
inhibition of an enzyme involved in its synthesis. After the product has been utilized or broken
down and its concentration thus decreased, the inhibition is relaxed, and the formation of the
product resumes. Such enzymes, whose ability to catalyze a reaction depends upon molecules
other than their substrates (the ones upon which they act to form a product), are said to be under
allosteric control. Feedback inhibition is a mechanism by which the concentration of certain cell
constituents is limited.