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 ..