enzimologi reg nt pkii
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enzimologiTRANSCRIPT
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ENZIMOLOGIENZIMOLOGI
NINA TRISTINANINA TRISTINASUB-BAGIAN KIMIA KLINIKSUB-BAGIAN KIMIA KLINIK
BAGIAN PATOLOGI KLINIK FKUP/RSHS BANDUNGBAGIAN PATOLOGI KLINIK FKUP/RSHS BANDUNG
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ENZIMOLOGI
ILMU YANG MEMPELAJARI ENZIM
ENZIM :
- PROTEIN
- KATALITIK PROTEIN
- KONFIGURASI 3D AKTIF
- SPESIFIK THD SUBSTRAT/ Rx KIMIA ttu.
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REAKSI :
E + S ES PRODUK + E
(BTK ANTARA)
SATUAN :
* UNIT (U, mU) :
x µmol substrat yg mampu diubah oleh E dalam
waktu 1 menit
* KATAL :
x mol substrat yg mampu diubah oleh E dlm wkt
1 detik (nkat, µkat)
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• AKTIVITAS DIPENGARUHI:
- SUHU (>40º C)
- ASAM/ BASA
- CHAOTROPIC AGENTS : Urea, protease
- ADDITIVE i.e EDTA
• VARIAN : ISOFORMS
- TRUE ISOENZYME : LOKUS BERBEDA ;
ALLELOZYMES (INHERITANCE);
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• KONVERSI
µ kat/L 60 U/L
nkat/L 0,06 U/L
U/L 0,0167 µ kat/L
U/L 16,67 nkat/L
• NAMA ENZ (Enz.Commission)
e.g CREATINE KINASE =
ATP: creatine N.phosphotransferase,
EC 2.7.3.2
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• KLASIFIKASI :
1. Oksidoreduktase
2. Transferase
3. Hidrolase
4. Lyase
5. Isomerase
6. Ligase
Kec. Chymotrypsin
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• KEPENTINGAN KLINIS :
- ENZ. TEST : HEPATO BILIER
- ENZ TEST : GASTROENTEROLOGI
- ENZ. TEST : KARDIOVASKULER
- ENZ. TEST : PENY. OTOT SKELET
- ENZ. TEST : PENY. TULANG
- ENZ. TEST : KEGANASAN
- ENZ. TEST : RENAL & TR. UR.
- ENZ. TEST : BODY FLUIDS
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• TIDAK ADA ENZ. YANG BENAR-BENAR
DIHASILKAN HANYA OLEH 1 ORGAN/ SEL
PANEL PEM. ENZIM
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• PERSIAPAN P/ & BP
- PUASA 8-10 JAM SEBELUMNYA
- SAMPLING : Pdu : SERUM
* JGN. HEMOLISIS
* SESEGERA MUNGKIN
* WADAH BP
• PANEL : KESEPAKATAN
• INTERPRETASI : SELALU BERPEDOMAN PD
KEADAAN KLINIS
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• ENZIM YANG SERING DIPERIKSA :
1. SGOT, SGPT
2. ALP, γGT
3. LDH
4. AMILASE, LIPASE
5. CK/CPK, CKMB
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• NILAI RUJUKAN/ REFERENSI:
- DARI POPULASI
- BELUM TENTU DISEBABKAN OLEH PENYAKIT BILA HASIL PEM. DI LUAR NILAI RUJUKAN
- TGT. SUHU PEMERIKSAAN & METODE
- PTG!! : KLINIS & HSL. PEM.
- PERLU DIINGAT ADANYA INTERFERING FACTORS
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• HAL- HAL KHUSUS :
- γ GT : PEM. SINGEL MENINGKAT ULANG PADA HARI YANG BERBEDA
- ENZ. JANTUNG TIDAK MENINGKAT, TETAPI KLINIS (+) AMBIL BP PADA JAM-JAM BERIKUTNYA
- KEADAAN KRONIS : PENINGKATAN MINIMAL e.g pd SIROSIS : KADANG-KADANG NORMAL KEGAGALAN SEL
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PRINCIPLES OF ENZYME ACTIVITY PRINCIPLES OF ENZYME ACTIVITY DETERMINATIONDETERMINATION
Enzyme concentration Enzyme concentration ~ enzyme activity~ enzyme activity
International Unit/ L (IU/ L)International Unit/ L (IU/ L)
Based on: enzyme – specific substrate reactionBased on: enzyme – specific substrate reaction kk11 k k33
E + S E + S (E S) (E S) E + P E + P kk22E : ENZYME E : ENZYME S : SUBSTRATES : SUBSTRATEES : ENZYME-SUBSTRATE COMPLEXES : ENZYME-SUBSTRATE COMPLEXP : PRODUCTP : PRODUCTK1 : forward rate constantK1 : forward rate constant
K2 : reverse rate constantK2 : reverse rate constantK3 : covalent catalytic constantK3 : covalent catalytic constant
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ENZYMES MEASUREMENTENZYMES MEASUREMENT
BIOCHEMISTRY ASSAYSBIOCHEMISTRY ASSAYS IMMUNOCHEMICAL ASSAYS IMMUNOCHEMICAL ASSAYS ISOENZYMESISOENZYMES ELECTROPHORETIC ASSAYELECTROPHORETIC ASSAY
PITFALLS IN ENZYME ASSAYS :PITFALLS IN ENZYME ASSAYS :1.1. HemolysisHemolysis2.2. Anticoagulants; Anticoagulants; serum is the preferred serum is the preferred
specimenspecimen3.3. pHpH4.4. Specimen storage & Assay temperaturesSpecimen storage & Assay temperatures5.5. Iso-enzymesIso-enzymes
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CLINICAL APPLICATION OF ENZYME CLINICAL APPLICATION OF ENZYME ASSAYSASSAYS
DiagnosisDiagnosis Monitoring the course of the diseaseMonitoring the course of the disease Demonstrating Demonstrating subclinical subclinical evidence of evidence of
diseasedisease
ImportantImportant : :
disease can be characterized by distinctly disease can be characterized by distinctly
abnormal values of abnormal values of one or moreone or more enzymes; enzymes;
panelpanel of of enzymeenzymess examination examination
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ACTION COULD BE :
1. ACTIVE IN PLASMA : - COAGULASE
2. ACTIVE IN EXTRACELLULAR FLUID:
- DIGESTIVE ENZYMES
3. ACTIVE IN INTRACELLULAR:
- METABOLIC ENZYMES
TEMPERATURE DIFFERENT ACTIVITY
PLASMA ENZYME
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1. DIGESTIVE DISORDERS
2. PANCREATITIS
3. HEPATITIS
4. MYOCARDIAL INFARCTION
5. BONE DISEASE
6. MUSCULAR DISORDERS
7. INTOXICATION
8. MALIGNANCY
9. ERYTHROCYTE ENZYME DEFICIENCY
10. VITAMIN DEFICIENCY
ENZYMES TEST IN CLINICAL SETTING