BIOLOGI MOLEKULER BIOSINTESIS PROTEIN Oleh BAGIAN BIOKIMIA FAKULTAS

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 nucleus nucleolus cell membrane mitochondria r. EPR (ribosomes) s. EPR centrosome cytoplasm Golgi

Fungsi organela sel organelle major functions cell membrane nucleus nucleolus centrosome cytoplasm mitochondria ribosomes lysosomes r. EPR s. EPR Golgi cell integrity, signal transduction replication, transcription ribosome synthesis (subunits) cell division metabolic pathways, signaling ATP synthesis, DNA protein synthesis intracellular degradation protein synthesis (export) lipid synthesis, membranogenesis protein transport (addressing)

Structure of mammalian genes REGULATION STRUCTURE I 1 5‘ REGULATION E 1 E - exons (coding) I - introns (non-coding) I 2 E 3 promoter enhancer silencer 3‘

Structural parameters of a gene and of a m. RNA (comparison) Gene - 5‘-flanking region, structural part, 3‘-flanking region - exons (coding) and introns (non-coding) - no real start, no real stop c. DNA (m. RNA) - 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 E 1 DNA E 2 transcription + capping E 3 poly A signal termination signals CAP pre-m. RNA polyadenylation AAAAAA m. RNA splicing start AAAAA stop

Jenis RNA ribosomal RNA (r. RNA) 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 (m. RNA) matrix for protein synthesis at the ribosome, different length translation

Struktur m. RNA gene I 1 E 1 5‘ I 2 E 3 E 2 3‘ PAS pre-m. RNA AUG (start) m. RNA splicing sites PAS CAP 5‘-UTR start UGA (stop) TR AAAAAAA 3‘-UTR stop

Struktur m. RNA jadi CAP regulatory parts structural part 5‘-UTR start codon 3‘-UTR AAAAA TR stop codon

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

Bahasa gen bersifat 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 m. RNA SEBAGAI PEMBAWA INFORMASI GENETIK SINTESIS PROTEIN TERJADI DI RIBOSOM m. RNA MEMBAWA CODON, SEDANGKAN t. RNAMEMBAWA ANTI CODON PESANAN ASAM AMINO DIKODE OLEH CODON m. RNA 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 m. RNA 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 polypeptide chain (a-helix, ß-sheet, unstructured turns) tertiary structure spatial arrangement of secondary structural elements quatery structure subunit interactions

Amino acids are constituents of proteins there 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 k. Da Koloni yg mengeksp protein rekombinan diimunobloting dg Ab poliklonal thd Prot sol T. gondii 97, 4 k. Da 84, 0 k. Da 66, 0 k. Da 55, 0 k. Da 45, 0 k. Da 29, 0 k. Da 20, 1 k. Da 14, 2 k. Da 1 2 3 4 5 6 7 Pita prot rek gb pita berbeda dg prot non rek, 28 intensitas warna berbeda, yg dominan prot rek dg BM 28 k. Da yg merup hasil eksp c. DNA Gambar 6. Hasil elektroforesis protein dari biakan E. coli rekombinan pembawa gen penyandi protein solubel 28 k. Da (GRA 2) 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( 2. 3. 4. 5. 6. Hormon) Biokatalisator ( Enzim ) Protein Transport: Hb, Alb, Lipoprotein, Transferin Protein Kontraktil: aktin dan miosin Protein Struktural: kolagen, tubulin, keratin, glikopr Protein Pelindung: Ig, interferon, perforin, IL

HEALTH is better than WEALTH Biomol: Dx dini, patogenesis, protein kandidat vaksin dan tx gen

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TERIMA KASIH