Biochemistry

1
Biochemistry Molecular Biology
2
Outline

• Biochemistry and function of macromolecules and
  their derivatives
• Structure,properties,function of amino acid and
  protein
• Energy metabolism
• Metabolic pathway of small molecule
• Biosynthesis and degradation of macromolecule
• Gene expression
• Genetic engineering technology

3
Carbohydrate

• Definition
• Polyhydroxylated aldehydes and ketones
• Structure

4
Carbohydrate

• Sources and function
• ??????????????? starch, glycogen
• ????????? cellulose, chitin, murein
• ???????????????? mucopolysaccharides
  (chondrotin sulfate)
• ???????????? ribose, deoxyribose
• ?? cardiac glycoside (digoxin)

5
Carbohydrate

• Classification
• Monosaccharide
• Oligosaccharide
• Polysaccharide

6
Monosaccharide

• ????????????????????????????????????????
• ????????
• 1. ?????? (triose) ?? C 3 ????? ??????????????
  ?????? ??????????????? ????????????????????
• 2. ?????? (tetrose) ?? C 4 ????? ?????? ????????
  ?????????????
• 3. ?????? (pentose) ?? C 5 ????? ?????? ?????
  ???????????????????????????????????????????
  ??????? ??????????
• 4. ?????? (hexose) ?? C 6 ?????
  ??????????????????????? ?????? ?????? ??????
  ???????? ??????????

7
Monosaccharide

• derivative
• 1. ????????? (sugar acid) ??? ????????????????????
  ??????? (carboxylic group) ????????????? ????
  ??????????? (gluconic acid) ,??? D-???????????
  (D-glucuronic acid)
• 2. ????????????? (deoxy sugar) ???
  ?????????????????????????????????1 ????
  ????????????? ????????????? ???? D-2-deoxyribose
• 3. ???????????? (amino sugar) ???
  ?????????????????????????????????? ????
  D-?????????? ???????????????????????????????????
  (hyaluronic acid) ?????????????????? ????
  ????????????? (erythromycin) ?????????????????????
  ???????
•   4. ??????????????? (sugar alcohol) ???
  ??????????????????????????????????????????????????
  ? ????????? ???? ??????? (xylitol)
  ??????????????????????????????????
• 5. ???????????? (sugar phosphate)
  ???????????????????????????????????????? ??????
  ???????????????????????????????????? ????
  ??????????? ????????????????????? ??????????????
  ??????????????? ???????????????

8
Oligosaccharide

• Disaccharide
• Maltose
• Sucrose
• lactose
• Oligosaccharide ?????????? 3 ????? ?????????????
  glycoprotein

9
Polysaccharide

• ????????
• Homopolysaccharidestarch,glycogen,cellulose
• Heteropolysaccharide glycoprotein,
  glycoaminoglycans
• ???????
• ????????????
• ?????????

10
Glycoaminoglycan

• ????????? mucopolysaccharide
• ??????? peptide peptidoglycan
• ??????? protein proteoglycan
• ???????
• Ground substance
• ???? cell ??? bacteria

11
Glycoprotein

• ??? protein ???? oligosaccharide ????????
• ???????
• Structural molecule bone matrix ,fibrin
• Transport molecules for vit,mineral,lipid
• Cell attachmentrecognition site
• Immunologic moluculeIg,complement
• Enzyme
• Lubricantsprotective agents
• Lectin (????????? agglutination)

12
CBH metabolism

• Digestion
• Mouthintestine
• Absorption
• Small intestine
• Lactase deficiency
• Pancreatitis
• Parotitis

13
CBH pathway (glucose)

• Glycolysis
• Gluconeogenesis
• Cori cycle
• Pentose phosphate pathway
• Glycogenolysis
• Glycogenesis

14
Glycolysis

• ?????? cytoplasm ??? cell
• ??? product ???? pyruvate
• Anaerobic condition
• (lactate dehydrogenase)
• Aerobic condition
• (pyruvate dehydrogenase)

15
Gluconeogenesis

• ?????? liver and kidney
• Substrate
• Lactate
• Pyruvate
• Glycerol
• Oxaloacetate

16
Cori cycle
muscle
lactate
glucose
liver
17
Pentose phosphate pathway

• ???? pathway ?????????? RBC
• G6PD
• ?? cell ?????????????????????? ribose 6 phosphate

18
GlycogenolysisVSGlycogenesis

• ??????????????? liver and muscle
• Regulation
• Glucagon
• Epinephrine
• Insulin

19
Lipid

• ??? ??????????????????????????????????????????????
  
• Classification
• Simple lipid natural fat,oil,wax
• Compound lipid phospholipid,glycolipid,sphingolip
  id
• Derived lipid monoacylglyceral,diacylglyceral
• Miscellaneous lipid steroid

20
Fatty acid

• ??? aliphatic carboxylic acid
• Classification
• Saturated fatty acid
• Unsaturated fatty acid
• Monounsaturated
• Polyunsaturatedtrans gt cis
• Eicosanoid prostanoid,leukotriene

21
Essential fatty acid

• Linoleic acid
• Linolenic acid
• Arachidonic acid
• ???? substrate ??? eicosanoid cascade

22
Major type of lipid in animal tissue

• Triglyceride
• Wax
• Phospholipid
• Sphingolipid
• Steroid

23
Triglyceride

• ???????
• Heat insulator
• Cushion
• Storage food
• ?????????????????????????????
• ???????????????

24
Phospholipid

• Amphipathic compound
• ???????
• Major component of cell membrane
• Surfactant (phosphatidylcholine)
• Major component of lipoprotein

25
Sphingolipid

• Major component of membrane lipids
• Subclassification
• Sphingomyelin
• Cerebrocide
• ganglioside

26
Steroid

• Cholesterol
• Temperature buffer of Cell membrane
• Bile acid
• Hormone
• Ergosterol
• ????????????????????????????? vit D ??????????
• Anti-imflammatory agent

27
Lipid metabolism

• Digestion
• Mouth Stomach
• Duodenum
• Small intestine
• Absorption
• Micelles
• Small and medium chain fatty acid, glycerol

28
Lipid circulation

• Chylomicron
• VLDL
• LDL
• HDL
• HMG CoA reductase

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Fatty acid metabolism

• ?????? mitochondrial matrix
• Carnitine
• ß-oxidation
• Ketogenesis ?????????? starvation ????????
  acetoacetate ?????????????? blood-brain barrier
• DKA

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Clinical relevance

• Bile duct obstruction
• Pancreatic duct obstruction
• Small intestine disease
• Hyperlipidemia

31
Protein

• ???????
• Enzyme
• Peptide hormone
• Toxin
• Structural protein
• Storage protein
• Protective protein
• Transport protein
• Contractile protein
• Coagulation factor

32
Essential amino acid
??????? amino acid 1.building block 2.cellular
signal (GABA) 3.neurotransmitter
(dopamine) 4.inflammatory mediator
(histamine) 5.hormone (thyroxine)

• Arginine
• Phenylalanine
• Isoleucine
• Tryptophan
• Threonine
• Lysine
• Leucine
• Valine
• Histidine
• Methionine

33
Protein

• Primary structure (linear)
• Secondary structure
• Alpha-helix
• Beta-plated sheet
• Tertiary structure
• Fibrous
• Globular
• Quaternary structure (subunit)

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Enzyme

• ????????????
• E S ES P E

35
Enzyme

• ????????????????????????
• ????
• ?????? enzyme ??? substrate
• First-order reaction
• Zero-order reaction
• Temperature
• pH

36
Term

• Km (??? substrate ?????? enzyme ?????
• Vmax (??? enzyme ??????????
• Enzyme inhibitor
• Isosteric VS allosteric inhibitor
• Irreversible inhibitor
• Reversible inhibitor Km Vmax
• Competitive inhibitor (AS) ? ?
• Non-competitive inhibitor (NS) ? ?
• Uncompetitive inhibitor (ES) ? ?

37
Protein matabolism

• Digestion
• Stomach
• Pancreatic juice
• Absorption
• Amino acid only

38
Amino acid metabolism

• Removal of amino acid nitrogen
• Transamination (AST,ALT)
• Deamination (glutamate dehydrogenase)
• Product ??? NH4 and carbon skeleton

39
Detoxification of NH4

• Peripheral tissue
• Skeletal muscle ?????? NH4 ?????? alanine
• Other tissue ?????? NH4 ?????? glutamine
• Liver
• ??????? amino acid ??????? NH4 ???????? urea
  cycle ??????????????????? urea ?????????????
  urine

40
Electron transport

• Complex I (NADH)
• Complex II (FADH2)
• Complex III
• Complex IV (ATP synthase complex)
• Inhibitor
• Complex I amobarbital rotenone
• Complex II antimycin A
• Complex III cyanide CO H2S

41
Nucleic acid

• Nucleoside
• basesugar
• Nucleotide
• Basesugarphosphate
• Nucleic acid
• Polymer (monomer ??? nucleotides)
• Genetic material

42
Nitrogenous base
Phosphate

• Purine
• Adenine
• Guanine
• Pyrimidine
• Thymine
• Cytosine
• Uracil

PO43-
Sugar

• Ribose
• Deoxyribose

43
Nucleotide

• ???????
• 1.???? building block ?????????? Nucleic acid
• 2.????????????????????? (ATP,AMP,GTP)
• 3.???? secondary massenger (cAMP)
• 4.???? coenzyme (NAD)
• 5.?????? (Adenosine , acido thymidine
  ,lamivudine)

44
DNA

• Chargaffs rule
• ??????????????? DNA ??? ???????????????????
• ??????????????? DNA ??? ????????????????????
• ??????????????? DNA ??? ????????????????????
• ?? DNA ?????????????????????? ????
  purinepyrimidine ???? (AT ,CG)
• ???????????? polydeoxyribonucleotide 2
  ???????????????? antiparallel ????? specific base
  pairing

45
RNA
Nucleoprotein

• Ribosomal RNA
• Messenger RNA
• Transfer RNA

• Nucleic acidprotein
• Chromosome
• Ribosome

46
Term

• Chromosome
• Nucleosome
• Histone
• Non histone protein

Clinical relevance

• Gout result from purine metabolism disorder

47
?????????????

• 5 ACUGGCCAACAGA 3

48
Gene expression
DNA
Replication
Transcription
Translation
RNA
Protein
49
Replication

• Helicases unwind the DNA duplex
• Primase makes an RNA primer (5 3)
• Leading strand
• Lagging strand ,Okazaki fragments
• DNAP I remove and replace RNA primer
• DNA ligase join the end together

50
Transcription

• Binding (specific promoter sequences)
• Initiation
• Elongation
• Termination (hairpin loop)
• After transcription process
• Cap at 5 end (7-methylguanine)
• Poly A tail at 3 end
• RNA splicing (remove intron)

51
Translation

• ????????? AUG
• ???????? UAA UAG UGA
• ???????? 5 ?? 3 ??? mRNA
• ?????????? ribosome

52
Mutation

• Substitution
• Transition
• Transversion
• Insertion or deletion
• Frameshift mutation

• Missense
• Nonsense
• Silent

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Genetic engineering technology

• Southern blot (DNA) (restriction endonuclease)
• Northern blot (RNA) (restriction endonuclease)
• Western blot (Protein) (antibody)
• PCR (DNAprimerdNTPDNAP)
• DNA fingerprint (polymorphisms) (restriction
  fragment length polymorphism)

54
Quiz
55

• 1. ??????????? 3 ?? ??????? ??????
  ??????????????????
• ??? kwarsiokor ??????????????? amino acid ??????
• ????
• a. Threonine
• b. Tryptophan
• c. Methionine
• d. Aspartic acid
• e. Phenyl alanine

56

• 2. ?????????????????? ????? ????????????????????
• ?????? ethanol ????????????????????
  ???????????????
• ??? ethanol ???
• ?. Product inhibitor
• ?. Allosteric inhibitor
• ?. Irreversible inhibitor
• ?. Competitive inhibitor
• ?. Feedback inhibitor

57

• 3. ??????????????? ????????????????? deep tendon
• reflex ????????? metabolic pathway ?????????
• a. glycolysis
• b. Krebs cycle
• c. Glucogenesis
• d. Glycogenesis
• e. Oxidative

58

• 4. ??????????????? acute lymphoblastic leukemia
  ??????
• ??????????? ?????? uric acid ??? metabolism ?????
• ???????
• a. heme
• b. purine
• c. pyrimidine
• d. amino acid
• e. glucoronide

59

• 5. ??????? 50 ?? ?????????????????? 10 ??
  ?????????
• epigastric ????????????? ??????????????
  ???????????? ??
• ???????????????????????
• a. stomach
• b. liver
• c. pancrease
• d. small intestine
• e. gallbladder

60

• 6. ????????????????????????????
  ?????????????????
• ???????
• a. micelle
• b. chylomicron
• c. cholesterol
• d. fatty acid
• e. monoglycerine

61

• 7. ??????? 50 ?? ????????????????????????????????
  ?????
• ???????????????? ????????????? ????????? ????????
• ?????????? ???????????????????? 5 ??.
  ?????????????
• ????? ??????? Hb ????
• a. sulfhemoglobin
• b. methamoglobin
• c. oxyhemoglobin
• d. carboxyhemoglobin
• e. oxidized hemoglobin

62

• 8. ????????????? bile salt ?????????????
  ??????????
• ?????????????
• a. bilirubin
• b. lecithin
• c. fatty acid
• d. triglyceride
• e. cholesterol

63

• 9. ????????????????????? ??????????????
  ?????????
• ????????????????????? ?????????????? ???????
• ????????????????? ???? VDRL ???????
  ??????????????
• ???????? 2 ??. ????????? Hb typing
  ???????????????
• a. alpha 4
• b. beta 4
• c. gamma 4
• d. delta 4
• e. zeta 4

64

• 10. ??????? 50 ?? ?????????????????
  ????????????????????????? 2 ???? ???
  ??????????????????????????????????????? Hb 8 g/dl
  Hct 24 WBC 6000 N75L25 platelet 200000 MCV 80
  fL reticulocyte 1creatinine 5 ???????????????????
  ????? ???????????????????????????
• a. Folic acid
• b. Erythropoietin
• c. Thyroid hormone
• d. Cyanocobalamine
• e. Colony stimulating factor

65

• 11. ??????????? 5 ?? ????????????????????????????
  ??
• ????????? ??????????????????? ?????????????
• platelet 200,000/ cu.mm. ???????????????????????
• ?????????????????????
• a. Deamination of alanine
• b. Hydroxylation of proline
• c. Phosphorylation of serine
• d. Glycosylation of hemoglobin
• e. Carboxylation of glutamic acid

66

• 12. ??????? 30 ?? ???? thalassemia
  ??????????????????????
• ????????? serum glucose 140 ???????????????????
• ???????????
• a. Insulinitis
• b. Pancreatitis
• c. ?? amyloid ????
• d. Hemochronatosis

67

• 13. N-acetylcystein????????????????????
• paracetalmol toxicity ????????????????????????????
  ?
• ????????? ??? liver
• a. N-acetylation
• b. N-hydroxylation
• c. sulfer conjugate
• d. glutathione conjugate
• e. glucolonide conjugate

68

• 14. ??????? 40?? ?????? first metatarsophalangeal
  
• joint , serum uric acid ??? ????????
• a. ??????? gout
• b. ???????????????????? pyrimidine salvage
• pathway
• c. ????????????? nucleic acid ???
• d. ?????? acidosis ???

69

• 15. ?????????????? B-thalassemia \ HbE disease
• ??????????????????? HbE homozygote ??????????????
• ??? thalassemia ????????????
• A. 1
• B. 3/4
• C. 1/2
• D. 1/4
• E. 0

70

• 16. Pathway ???????????????? RBC
• a. Urea cycle
• b. Krebs Cycle
• c. Beta oxidation
• d. Lactose fermentation
• e. Glycolysis

71

• 17. ?????????????? 60 ?? ????????????????????????
  ???
• ??????? ????????????? ???????????????????? 1 ??.
  ???
• ?????????? ??????????????????????????????????
• a. stercobillin
• b. serum albumin
• c. urine urobillinogen
• d. plasma prothrombin
• e. serum direct bilirubin

72

• 18. ??????? 40 ?? ?????????????????????????
  ?????
• ???????????? 2 ?? ????????????????????? ?? 4 ??
  ?????
• ??????????????????????????????
• a. Nose
• b. nasopharynx
• c. Trachea
• d.bronchus
• e. alveoli

73

• 19. ??? 45 ?? ???????????? ??? MTP jt.
  ?????????????
• ??????????? uric 8.2 mg/dl ??????? metabolism ???
• ?????
• a. purine
• b. protein
• c. glucose
• d. cholesterol
• e. pyrimidine

74

• 20. ??????????????????????? mutatuion ????
• ..ATGCCAAA.. ???????????? DNA probe ???????????
• ?????
• ?. UUUGGCAU
• ?. UACGGUUU
• ?. TTTGGCAT
• ?. TACGGTTT
• ?. TTTGGUAT

75

• 21. ??????????????? 35 ?? ???????????????????????
  ????
• ??????????????????????? ??????????????????????????
  ?????
• ???? ???????????????????????? single treatment
  ???
• ??????????
• 1. Ca
• 2. Vit D
• 3. Prednisolone
• 4. Progesterone
• 5. Estrogen

76

• 22. ????????? ????????? Hb H ??????????????????
• Hct Hb MCV Reticulocyte
• ???? 38 13 72 1
• ????? 39 13 87 0.5
• Genotype ???????????????
• a. ???? aa/--
• b. ????? a-/a-
• c. ????? aa/aa
• d. ???? ßoßt
• e. ????? ßoßo

77

• 23. ???????????? 2 ?? ??? Vit ????
• 1. thiamine
• 2. folate
• 3. Riboflavin
• 4. B12
• 5. niacin

78

• 24. ???????? 35 ?? ??? ??????? ??????????? ????
• peptic ulcer ????????????? ???????????????????
• ???? ????? ????????? ???????? ???????? ??? ??????
• ??????????? Hb 8 g/dL Hct 30 Reticulocyte 8
• Platelet 250,000/mm3 MCV 86 fL ????????????????
• ???????????
• 1. autoimmune
• 2. ??? Vit B12
• 3. Hb ???????
• 4. ??? enzyme
• 5. Post hemorrhage

79

• 25. ??????? 5 ?? ????????????????? down syndrome
• ????????? karyotype ??? 46,XY ????????????????????
  
• ????????????
• 1.deletion
• 2.mosaicsm
• 3.translocation
• 4.somatic mutation
• 5.undetected trisomy

80

• 26. ???? 25 ?? ?????????? ????????????????
  ???????
• ????????????? ???????????????????????????
• ??????? ????????????????????????
• 1. paraquat
• 2. organophosphate
• 3. ethylene glycol
• 4. arsenical compound
• 5. sodium hypochlorite

81

• 27. lipid ?????????????????????????? a.
  cholesterol
• b. sphingolipid
• c. phospholipids
• d. fatty acyl
• e. triglyceride

82

• 28. ??????? 25 ?? ???????????? potassium cyanide
  ????? ?????????????????????????????? ??????
  cyanide poisoning ??????????????????
• a. glycolysis
• b. Krebs
• c. Electron transport chain
• d. phosphooxidative

83

• 29. ????????? Paraquat ???? superoxide
  ???????????? cell injury??????? ??????
  ????????????????? O2 free radical)
• a. B1 Thiamine
• b. B6
• c. A (Retinoic acid)
• d. C (Ascorbic acid)
• e. B5 Panthetoic acid

84

• 30. - - - GCSCGG - - -???? - - - GCCGG- - -
  ????????????????????
• a. non sense
• b. missense
• c. frame shift
• d. silence

85

• 31. ??????? ???? 25 ?? ?????????????????????
  ????????? ?? 3 ??? CBC Hb 10 WBC 7,000 Plt
  300,000 Reticulocyte 7 MCV 92 ??????????????
• a. iron def
• b. B12 def
• c. Hemolytic anemia
• d. Hepoglobinopathy
• e. Post hemorrahagic anemia

86

• 32.?????? ???? 50 ?? ???? gout ???????????
  allopurinol ???????????????????? enz ??
• 1.xanthine oxidase
• 2.adenine deaminase
• 3.guanine deaminase
• 4.adenine phosphoribosyltransferase
• 5.guanine phosphoribosyltransferase

87

• 33.Carnitine ?????????????????????????????????????
  ?
• 1.glucose
• 2.glycogen
• 3.amino acid
• 4.fatty acid
• 5.triglyceride

88

• 34.Metabolism ??? CBH 100 gm ,protein 20g,lipid
  25 g ????????????????
• 1.300 kcal
• 2.500 kcal
• 3.700 kcal
• 4.900 kcal
• 5.1100 kcal

89

• 35. ??????? 5 ?? ????????????????? down syndrome
• ????????? karyotype ??? 46,XY ????????????????????
  
• ????????????
• 1.deletion
• 2.mosaicsm
• 3.translocation
• 4.somatic mutation
• 5.undetected trisomy