Enzimologi Reg NT PKII

ENZIMOLOGIENZIMOLOGI

NINA TRISTINANINA TRISTINASUB-BAGIAN KIMIA KLINIKSUB-BAGIAN KIMIA KLINIK

BAGIAN PATOLOGI KLINIK FKUP/RSHS BANDUNGBAGIAN PATOLOGI KLINIK FKUP/RSHS BANDUNG

ENZIMOLOGI

ILMU YANG MEMPELAJARI ENZIM

ENZIM :

- PROTEIN

- KATALITIK PROTEIN

- KONFIGURASI 3D AKTIF

- SPESIFIK THD SUBSTRAT/ Rx KIMIA ttu.

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)

• AKTIVITAS DIPENGARUHI:

- SUHU (>40º C)

- ASAM/ BASA

- CHAOTROPIC AGENTS : Urea, protease

- ADDITIVE i.e EDTA

• VARIAN : ISOFORMS

- TRUE ISOENZYME : LOKUS BERBEDA ;

ALLELOZYMES (INHERITANCE);

• 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

• KLASIFIKASI :

1. Oksidoreduktase

2. Transferase

3. Hidrolase

4. Lyase

5. Isomerase

6. Ligase

Kec. Chymotrypsin

• 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

• TIDAK ADA ENZ. YANG BENAR-BENAR

DIHASILKAN HANYA OLEH 1 ORGAN/ SEL

PANEL PEM. ENZIM

• 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

• ENZIM YANG SERING DIPERIKSA :

1. SGOT, SGPT

2. ALP, γGT

3. LDH

4. AMILASE, LIPASE

5. CK/CPK, CKMB

• 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

• 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

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

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

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

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

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