REGULASI AKTIVITAS ENZIM

aturan sistem yang ada di dalam tubuh makhluk hidup untuk dapat hidup seimbang, mempertahankan keadaan teratur, konservasi energi, dan sebagai respon terhadap perubahan lingkungan

REGULASI AKTIVITAS ENZIM

Berbagai reaksi enzimatis tidaklah berjalan secara mekanis begitu saja, tanpa ada pengendalian Reaksi enzimatis berjalan secara sangat terkoordinasi satu sama lain Pengendalian reaksi enzimatis berlangsung pada berbagai tingkat di dalam sel, semenjak dari gen sampai molekul enzim itu sendiri.

 Regulasi aktivitas enzim dilakukan oleh:

Pengendalian pada tingkat Gen

Pengendalian ditingkat molekul enzim oleh produk Pengendalian enzim melalui perubahan struktur molekul

Pengendalian pada tingkat gen

Sebagai protein, informasi genetik enzim terekam dalam gen Sel hanya akan mensintesis suatu enzim, jika sel mengandung gen yang menyandikan enzim yang dimaksud

So how can genes be turned off?

Langkah pertama pembuatan protein
transcription stop RNA polymerase!

Repressor protein
binds to DNA near promoter region blocking RNA polymerase
binds to operator site on DNA blocks transcription

Pengendalian pada tingkat gen

Terdapat 2 mekanisme pengaturan secara genetik

Inducible enzyme (enzim terbangkitkan)

Represi enzyme (pembungkaman enzim)

Enzim konstitutif

Repressor protein model

RNA polymerase

Operon: operator, promoter & genes they control serve as a model for gene regulation

RNA repressor TATA polymerase

gene1

gene2

gene3

gene4

DNA

promoter

operator

Repressor protein turns off gene by blocking RNA polymerase binding site.

repressor

repressor protein

Repressible operon: tryptophan

Synthesis pathway model
RNA polymerase

When excess tryptophan is present, binds to tryp repressor protein & triggers repressor to bind to DNA
blocks (represses) transcription

RNA repressor TATA polymerase

gene1

gene2

gene3

gene4

DNA

repressor

repressor protein tryptophan (corepressor)

promoter

operator
tryptophan repressor protein complex conformational change in repressor protein!

repressor

Inducible operon: lactose

Digestive pathway model
RNA polymerase

When lactose is present, binds to lac repressor protein & triggers repressor to release DNA
induces transcription

RNA repressor TATA polymerase

gene1

gene2

gene3

gene4

DNA

repressor

repressor protein lactose (inducer)

promoter

operator
lactose repressor protein complex conformational change in repressor protein!

repressor

Operon summary
Repressible operon
usually functions in anabolic pathways
synthesizing end products

when end product is present in excess, cell allocates resources to other uses

Inducible operon
usually functions in catabolic pathways,
digesting nutrients to simpler molecules

produce enzymes only when nutrient is available

cell avoids making proteins that have nothing to do, cell allocates resources to other uses

The BIG Questions

How are genes turned on & off in eukaryotes? How do cells with the same genes differentiate to perform completely different, specialized functions?

Why turn genes on & off?

Specialization
each cell of a multicellular eukaryote expresses only a small fraction of its genes

Development
different genes needed at different points in life cycle of an organism
afterwards need to be turned off permanently

Responding to organisms needs

homeostasis cells of multicellular organisms must continually turn certain genes on & off in response to signals from their external & internal environment

Points of control
The control of gene expression can occur at any step in the pathway from gene to functional protein
unpacking DNA transcription mRNA processing mRNA transport
out of nucleus through cytoplasm protection from degradation

translation protein processing protein degradation

 Control regions on DNA

promoter

Transcription control : initiation step

nearby control sequence on DNA binding of RNA polymerase & transcription factors base rate of transcription

enhancers
distant control sequences on DNA binding of activator proteins enhanced rate (high level) of transcription

Model for Enhancer action

Enhancer DNA sequences

distant control sequences

Activator proteins
bind to enhancer sequence & stimulates transcription

Silencer proteins
bind to enhancer sequence & block gene transcription

Translation control
Block initiation stage
regulatory proteins attach to 5 end of mRNA
prevent attachment of ribosomal subunits & initiator tRNA block translation of mRNA to protein

 Protein processing

Protein processing & degradation control

folding, cleaving, adding sugar groups, targeting for transport

Protein degradation
ubiquitin tagging proteosome degradation

Ubiquitin
Death tag

1980s | 2004

mark unwanted proteins with a label 76 amino acid polypeptide, ubiquitin labeled proteins are broken down rapidly in "waste disposers"
proteasomes

Aaron Ciechanover Israel

Avram Hershko Israel

Irwin Rose UC Riverside

Proteasome
Protein-degrading machine
cells waste disposer can breakdown all proteins into 7-9 amino acid fragments

 Pengendalian ditingkat molekul enzim oleh produk

Pengendalian dilakukan melalui Alosterik kontrol atau feedback inhibition. Regulasi alosterik mengatur aktivitas enzim melalui hasil akhir (end product). Pada sistem ini End product akan menghambat pembentukan enzim pertama (enzim alosterik) yang mengawali jalur reaksi tersebut bilamana hasil akhir melebihi kebutuhan sel

Alosterik kontrol

Reminder: Regulation of metabolism

Feedback inhibition product acts as an allosteric inhibitor of 1st enzyme in tryptophan pathway

= inhibition

Alosterik kontrol atau Feedback control dibedakan atas :

Simple feedback
Concerted feedback Multiple enzim feedback Commulative feedback

feedback inhibition
Figure 3. Jalur biosintesis tryptophan pada E. coli. Jalur ini diregulasi oleh proses feedback inhibition.

Figure 4. Skema umum regulasi jalur metabolisme bercabang oleh proses feedback inhibition.

1. Simple feedback

2. Concerted feedback 3. Multiple enzim feedba 4. Commulative feedback

Another way to regulate metabolism

Gene regulation
block transcription of genes for all enzymes in tryptophan pathway
saves energy by not wasting it on unnecessary protein synthesis

= inhibition

Cascade Amplification of Signals

Pengendalian enzim melalui perubahan struktur molekul

Seringkali juga keadaan memerlukan sel untuk mengolah substrat dalam waktu yang singkat, melalui jalur metabolisme yang tidak selalu aktif. Untuk memenuhi kebutuhan seperti ini, enzim tertentu dibuat tidak aktif. Baru jika keadaan memerlukan enzim seperti ini diaktifkan melalui perubahan kimia yang ringkas, stabil namun cepat. Pengaktifan ini melalui perubahan struktur atau perubahan kovalen molekul enzim. Peristiwa ini sering dinamai modifikasi kovalen Modifikasi kovalen: searah (jika sudah aktif tidak bisa diinaktifkan), berbalik (tidak aktif aktif)

KOFAKTOR
Enzim terdiri atas 1 atau lebih rantai polipeptia, selain itu terdapat pula bagian yang bukan protein yang penting untuk aktivitas katalitik. Bagian yang bukan protein ini disebut KOFAKTOR. Kofaktor dibagi menjadi:

Gugus prostetik

Koenzim

ion metal

Beberapa contoh enzim yang memerlukan kofaktor

Aisjah Girindra. 1990. Biokimia I. Penerbit PT. Gramedia Jakarta. Hal. 91-113 Murray RK, dkk. 2003. Biokimia Harper. Penerbit Buku Kedokteran, EGC. Hal. 70-113 McGilvery dan Goldstein. 1996. Biokimia suatu pendekatan fungsional. Airlangga University Press. Hal. 299-339 Sadikin M. 2002. Biokimia Enzim. Widya Medika. 379 halaman Meisenberg, G dan Simmons WH. 2006. Principles of Medical Biocemistry. Hal. 43-68. Elsivier Mosby