BIOLOGI MOLEKULER

BIOSINTESIS PROTEIN
Oleh:

BAGIAN BIOKIMIA
FAKULTAS KEDOKTERAN UNS
2008
MAKNA SINTESIS PROTEIN
• MEKANISME ANTIBIOTIK DALAM
MEMBUNUH BAKTERI DENGAN
MENGHAMBAT SINTESIS PROTEIN
DINDING BAKTERI
• ANTIBIOTIK ADA YANG BERSIFAT
SELEKTIF MENGHAMBAT SINTESIS
PROTEIN DINDING BAKTERI SAJA,
TETAPI ADA YG MENGHAMBAT
KEDUANYA
Dogma of molecular biology

Produk akhir gen adalah protein

 Structure of eucaryotic cells

lysosomes cell membrane

mitochondria
nucleus

nucleolus rEPR (ribosomes)

sEPR
centrosome

cytoplasm Golgi
 Fungsi organela sel
organelle major functions
cell membrane cell integrity, signal transduction
nucleus replication, transcription
nucleolus ribosome synthesis (subunits)
centrosome cell division
cytoplasm metabolic pathways, signaling
mitochondria ATP synthesis, DNA
ribosomes protein synthesis
lysosomes intracellular degradation
rEPR protein synthesis (export)
sEPR lipid synthesis, membranogenesis
Golgi protein transport (addressing)
 Structure of mammalian genes
REGUL- REGUL-
STRUCTURE
ATION ATION
I1 I2
E1 E2 E3
5‘ 3‘

promoter
E - exons (coding)
enhancer
I - introns (non-coding)

silencer
Structural parameters of a gene
and of a mRNA (comparison)
Gene
- 5‘-flanking region, structural part, 3‘-flanking region
- exons (coding) and introns (non-coding)
- no real start, no real stop

cDNA (mRNA)
- 5‘-UTR, coding region, 3‘-UTR
- no uncoding region between start and stop
- start (cap-site) and stop (poly-A tail)
 Processes involved in gene
transcription
start stop
DNA E1 E2 E3
poly A termination
signal signals
transcription + capping
CAP

pre-mRNA polyadenylation
AAAAAA

splicing
mRNA
AAAAA
start stop
 Jenis RNA
ribosomal RNA (rRNA)
5 S, 5.8 S (160 bp), 18 S (2000 bp) and 28 S (5000 bp) RNA

ribosome structure

transfer RNA (t-RNA)

small RNA species with characteristic secondary structure
(anticodon, amino acid binding)

amino acid transfer

messenger RNA (mRNA)

matrix for protein synthesis at the ribosome, different
length
translation
 Struktur mRNA
gene
I1 I2
E1 E2 E3
5‘ 3‘

PAS
pre-mRNA
AUG UGA
(start) splicing sites (stop)

mRNA CAP PAS

AAAAAAA
5‘-UTR TR 3‘-UTR
start stop
 Struktur mRNA jadi

regulatory parts
structural part
CAP

5‘-UTR 3‘-UTR AAAAA

start TR stop
codon codon
 Genetic code
degenerated code: several triplets code for one amino acid,
only methionine (start) has just 1 codon

stop stop
stop
Bahasa
Bahasagengen
bersifat
bersifat
universal
universal
KODON ATAU SANDI GENETIK
TRIPEL NUKLEOTIDA
 LIHAT VCD SINTESIS
PROTEIN
• DARI MCB
• DARI SI NTESIS PROTEIN LEHNINGER 2000
• DARI HRN YAHYA
 BIOSINTESIS PROTEIN
• PERHATIKAN TEORI DOGMA SENTRAL, PRODUK
AKHIR GEN ADALAH PROTEIN
• DNA SEBAGAI MATERI GENETIK
• mRNA SEBAGAI PEMBAWA INFORMASI GENETIK
• SINTESIS PROTEIN TERJADI DI RIBOSOM
• mRNA MEMBAWA CODON, SEDANGKAN tRNA-
MEMBAWA ANTI CODON
• PESANAN ASAM AMINO DIKODE OLEH CODON mRNA
• ASAM AMINO YANG SATU DENGAN YANG LAIN
DIRANGKAI DENGAN IKATAN PEPTIDA
• START CODON SELALU DIMULAI DENGAN METIONIN
DAN SINTESIS PROTEIN DIAKHIRI DENGAN STOP
CODON
TABEL KODON
 Amplification of Message by Translation

From 1 spliced mRNA molecule many protein molecules are made.

SINTESIS PROTEIN SELALU DIMULAI
DENGAN METIONIN
TERMINASI DENGAN STOP CODON
 Structure levels in proteins
primary structure
amino acid sequence

secondary structure
spatial arrangement of amino acids within a poly-
peptide chain (helix, ß-sheet, unstructured turns)

tertiary structure
spatial arrangement of secondary structural elements

quatery structure
subunit interactions
Amino acids are constituents of
proteins

there are about 20 proteinogenic amino acids

 Proteins are Made of AAs
Connected by Peptide Bonds

• Ikatan peptida bersifat ikatan rangkap

spt 2 lembar kertas yang dilem (SRTLH)
arti dipeptida, tripeptida, jml AA & ikt pep
• Pada praktikum: berwarna ungu
 IMUNOBLOTING
kDa kDa Koloni yg mengeksp
protein rekombinan
97,4 kDa diimunobloting dg Ab
84,0 kDa poliklonal thd Prot sol
66,0 kDa
55,0 kDa T. gondii
45,0 kDa Pita prot rek gb pita
29,0 kDa berbeda dg prot non rek,
28
20,1 kDa intensitas warna
berbeda,yg dominan prot
14,2 kDa
rek dg BM 28 kDa yg
1 2 3 4 5 6 7 merup hasil eksp cDNA
Gambar 6. Hasil elektroforesis protein dari biakan E.coli rekombinan pembawa
gen penyandi protein solubel 28 kDa (GRA2) T. gondii yang diimunobloting dengan
antibodi poliklonal terhadap protein solubel takizoit T. gondii.
1. protein standar, 2. lisat protein dari E.coli dengan pewarnaan amidoblack,
4, 5, 6 dan 7 adalah hasil imunobloting protein dari klon rekombinan, sedang
3 adalah protein dari klon berwarna biru ( bukan rekombinan)
 APLIKASI KLINIS
• WESTERN BLOT UNTUK DIAGNOSIS
HIV
• STRUKTUR PROTEIN
Proteins Form Complex 20 Structures

• R Groups define the

behavior of an amino
acid and the structure
a protein domain folds
into.

• Acidic
• Basic
• Neutral
• Hydrophobic
• Hydrophilic
• Electrostatic
 MACAM DAN FUNGSI
PROTEIN
1. Protein Regulator /
Bioregulator( Hormon)
2. Biokatalisator ( Enzim )
3. Protein Transport: Hb, Alb,
Lipoprotein,Transferin
4. Protein Kontraktil: aktin dan miosin
5. Protein Struktural:
kolagen,tubulin,keratin,glikopr
6. Protein Pelindung: Ig, interferon,
perforin,IL
 HEALTH
HEALTH isisbetter
better
than
thanWEALTH
WEALTH

Biomol:
Biomol:Dx
Dxdini,
dini,patogenesis,
patogenesis,
protein
proteinkandidat
kandidatvaksin
vaksindan
dantx
txgen
gen
Thank you for
your kind
attentions ..
TERIMA KASIH