ASAM AMINO, PEPTIDA,
DAN PROTEIN
INDAH PURNAMA SARY
PENDAHULUAN
Protein → polimer alami yang terdiri dari
sejumlah unit asam amino yang berikatan amida
(peptida) satu sama lainnya
Asam amino → monomer protein
Peptida → oligomer dari asam amino
Telur, rambut, jaring labah, hemoglobin
ASAM AMINO
ASAM AMINO
Struktur umum :
dipolar
Asam amino α
Amfoterik :
ion zwitter
Jenis Asam Amino
O
Non Polar netral H2N CH C OH
Polar netral O
CH2
Polar Asam H2N CH C OH
CH2
Polar Basa CH2
lisina
C O
CH2
O OH
CH2 asam glutamat
H2N CH C OH CH2
O
CH CH3 NH2
H2N CH C OH
CH3
valina CH2
serina
OH
Latihan
Sebutkan jenis asam amino berikut berdasarkan
kepolarannya !
CO2-
S CO2- CO2-
NH3+
NH3+ NH2+
Methione
(Met; M) Phenylalanine Proline
(Phe; F) (Pro; P)
CO2- CO2-
HO
NH3+ NH3+
Serine Threonine
(Ser; S) (Thr; T)
Sebutkan jenis asam amino berikut berdasarkan
keasamannya !
CO2-
CO2-
HS
NH3+
NH3+
Cysteine HO
(Cys; C) Tyrosine
(Tyr; Y)
CO2-
N +
H3N CO2-
NH3+
N
H NH3+
Lysine
Histadine (Lys; K)
(His; H)
pH Isoelektrik
At some intermediate pH called the pI (isoelectric point),
the concentration of the dipolar ion is at a maximum and
the concentrations of anionic and cationic forms are
equal
Sintesis Asam Amino
Dari -Halo Carboxylic Acids
Melalui Strecker Reaction
Gabriel synthesis
Hidrolisis protein dalam sistem
kehidupan
-Halo Carboxylic Acids
O O
H2O
– +
CH3CHCOH + 2NH3 CH3CHCO NH4Br
Br +NH3
Strecker Reaction
O H2 N CN
C NH3 HCN C
H R H R
-aminonitrile
Heat H3O+, H2O
H3N COO
C
H R
Mekanisme reaksi Strecker Reaction
Gabriel Synthesis
Reaksi pada Asam Amino
Asilasi Esterifikasi
O O
+ –
CH3COCCH3 + H3NCH2COO + CH3CH2OH
HCl
O
O O +
–
Cl H3NCH2COCH2CH3
CH3CNHCH2COH
Peptida
Hidrolisis Peptida
Hidrolisis asam memutus semua ikatan peptida
Hidrolisis Enzimatis memutus ikatan peptida tertentu
menghasilkan fragmen yang bisa dipisahkan dan diidentifikasi
Enzim yang mengkatalisis pemecahan ikatan peptida adalah
protease, peptidase, dan enzim proteolitik
Enzim yang spesifik memotong ikatan peptida pada ujung-C :
Trypsin lysine dan arginine (lys dan arg)
Chymotrypsin Phenilalanine, tyrosine, tryptophan (phe, tyr,
trp)
Carboxipeptidase Ujung-C terminal
Sianogen bromida methionin (met)
Enzim yang spesifik memotong ikatan peptida pada ujung-N :
Pepsin Phenilalanine, tyrosine, tryptophan (phe, tyr, trp)
Termolisin ile, leu, val
Analisis Rantai Ujung Peptida
C terminal → carboxypeptidase
N terminal → Sanger’s methode
Edmand degradation
Sanger N-Terminal Analysis
- The N-terminal end of the polypeptide is labeled with 2,4-
dinitrofluorobenzene and the polypeptide is hydrolyzed
- The labeled N-terminal amino acid is separated from the
mixture and identified
Edmand Degradation
- Edman degradation involve
sequential cleavage and
identification of N-terminal amino acids
- Edman degradation works well for
polypeptide sequence analyses up to
approximately 60 amino acid residues
The N-terminal residue of the
polypeptide reacts with phenyl
isothiocyanate
The resulting
phenylthiocarbamyl derivative is
cleaved from the peptide chain
The unstable product rearranges
to a stable phenylthiohydantoin
(PTH) which is purified by HPLC
and identified by comparison
with PTH standards
Logika Penetapan Urutan
Insulin merupakan peptida dg 30 asam amino. Mula2 dihidrolisis dg
sempurna, dianalisis dan diperoleh rumusnya:
Ala2ArgAsnCys2GlnGlu2Gly2His2Leu4LysPhe3ProSerThrTyr2Val3
Dg metode Sanger, diketahui bahwa asam amino ujung N adalah Phe,
sedangkan dg karboksipeptidase diketahui ujung C adl Ala. Dg
kimotripsin diperoleh 3 fragmen, 2 Phe, dan 1 Tyr. Setelah dipisahkan,
ketiga fragmen asam amino tersebut dianalisis dg degradasi Edman
diperoleh :
A. Leu-Val-Cys-Gly-Glu-Arg-Gly-Phe
B. Val-Asn-Gln-His-Leu-Cys-Gly-Ser-His-Leu-Val-Glu-Ala-Leu-Tyr
C. Thr-Pro-Lys-Ala
Dg tripsin diperoleh Ala, dan 2 peptida. Peptida yg lebih pendek dianalisis
dg metode Edman, diperoleh:
D. Gly-Phe-Phe-Tyr-Thr-Pro-Lys
Dg logika maka dpt ditetapkan urutan insulin:
Phe-Val-Asn-Gln-His-Leu-Cys-Gly-Ser-His-Leu-Val-Glu-Ala-Leu-Tyr-Leu-
Val-Cys-Gly-Glu-Arg-Gly-Phe-Phe-Tyr-Thr-Pro-Lys-Ala
Latihan
1. Diberikan peptida dg urutan:
Ala-Gly-Tyr-Trp-Ser-Lys-Gly-Leu-Met-Gly
Tentukan fragmen yg terbentuk jk dihidrolisis dg:
a. Tripsin b. Kimotripsin c. Sianogen bromida
2. Bradikinin, suatu nonapeptida,
Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg
Tentukan fragmen yg terbentuk jk dihidrolisis dg:
a. Tripsin b. Kimotripsin c. Pepsin
3. Suatu peptida dihidrolisis dengan enzim tripsin
menghasilkan fragmen
- Hys-Lys
- Trp-Arg
- Ala-Val-Phe-Gly-Lys
Dihidrolisis dengan enzim chymotrypsin dihasilkan
fragmen
- Gly-Lys-Trp
- Ala-Val-Phe
- Arg-Hys-Lys
Dari degradasi Edmand didapatkan residu Ala dan
hidrolisis carboxypeptidase didapatkan fragmen Lys.
Tuliskan rantai peptida yang dimaksud !
Sintesis Peptida
Dg perlindungan ganda
R1 R1
perlindungan
asam amino H
H2N CH COOH P1 N CH COOH
aa1
R2 R2 O
perlindungan
karboksil
H2N CH COOH H2N CH C P2
aa2
R1 R2 O
H -H2O
P1 N CH COOH + H2N CH C P2
R1 O R2 O
H H
P1 N CH C N CH C P2
Sintesis fase padat (solid phase synthesis)
CH2 CH2 CH2 CH2
CH CH CH CH
O O
CH2
H2NCHC NHCHCO
R' R
Protein
Primary structure = the amino acid sequence
plus disulfide links
Secondary structure = conformational
acids eg : helix, pleated sheet
relationship between nearest neighbor amino
Tertiary structure : Refers to overall shape (how
the chain is folded) eg : Globular proteins
Quaternary Structure : Some proteins are
assemblies of two or more chains. The way in
which these chains are organized is called the
quaternary structure.
Hemoglobin, for example, consists of 4 subunits