BIOSINTESIS PROTEIN
mitochondria
cell membranelysosomes
Golgi
rEPR (ribosomes)
sEPR
centrosome
nucleolus
nucleus
cytoplasm
Organelle Major functions
Cell membrane Cell integrity, signal transduction
Nucleus Replication, transcription
centrosome Ribosome synthesis (sub units)
cytoplasm Metabolic pathways, signaling
mitochondria ATP synthesis, DNA
ribosomes Protein synthesis
Lysosomes Intra celullar degradation
rEPR Protein synthesis (export)
sEPR Lidpid synthesis, membranogenesis
Golgi Protein transport (addresing)
I1 I2
I - introns (non-coding)
promoter
enhancer
silencer
STRUCTUREREGULATI
ON
REGULATION
E1 E2 E3
E - exons (coding)
5‘ 3‘
Gene
cDNA (mRNA)
- 5‘-flanking region, structural part, 3‘-flanking region
- exons (coding) and introns (non-coding)
- no real start, no real stop
- 5‘-UTR, coding region, 3‘-UTR
- no uncoding region between start and stop
- start (cap-site) and stop (poly-A tail)
pre-mRNA
mRNA
DNA E1 E2 E3
transcription + capping
polyadenylationAAAAAA
splicingAAAAA
start stop
terminationsignals
poly Asignal
CAP
start stop
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
AUG
(start)
UGA
(stop)
pre-mRNA
5‘ 3‘E1 E2 E3
gene
I1 I2
PAS
splicing sites
mRNA PAS
start stop
5‘-UTR 3‘-UTR
AAAAAAA
CAP
TR
AAAAA
CA
P
TRstartcodon
stopcodon
5‘-UTR 3‘-UTR
structural part
regulatory parts
degenerated code: several triplets code
for one amino acid,
only methionine (start) has
just 1 codon
stop
stop
stop
Bahasa gen bersifatuniversal
KODON ATAU SANDI GENETIKTRIPEL NUKLEOTIDA
DARI MCB DARI SI NTESIS PROTEIN LEHNINGER 2000 DARI HRN YAHYA
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
primary structure
secondary structure
tertiary structure
quatery structure
amino acid sequence
spatial arrangement of amino acids
within a poly-
peptide chain (a-helix, ß-sheet,
unstructured turns)
spatial arrangement of
secondary structural elements
subunit interactions
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
I M U N O B L O T I N G
1 2 3 4 5 6 7
66,0 kDa
2820,1 kDa
45,0 kDa
29,0 kDa
14,2 kDa
55,0 kDa
84,0 kDa
97,4 kDa
Gambar 6. Hasil elektroforesis protein dari biakan E.coli rekombinan pembawagen penyandi protein solubel 28 kDa (GRA2) T. gondii yang diimunobloting denganantibodi 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, sedang3 adalah protein dari klon berwarna biru ( bukan rekombinan)
Koloni yg mengeksp protein rekombinan diimunobloting dg Ab poliklonal thd Prot sol T. gondii
Pita prot rek gb pita berbeda dg prot non rek, intensitas warna berbeda,yg dominan prot rek dg BM 28 kDa yg merup hasil eksp cDNA
kDakDa
• STRUKTUR PROTEIN
Proteins Form Complex 20 Structures
• R Groups define the behavior of an aminoacid and the structurea protein domain foldsinto.
• 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
TERIMA KASIH
Thank you for your kind attentions ..
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