The document discusses enzymology and enzymes. It provides definitions of key terms related to enzymes like cofactors, coenzymes, isoenzymes, and ribozymes. It describes the structure of enzymes including their active sites and discusses factors that influence enzyme activity like temperature, pH and inhibitors. It also mentions the diagnostic and therapeutic applications of enzymes.
Enzymes are biological molecules (typically proteins) that significantly speed up the rate of virtually all of the chemical reactions that take place within cells. They are vital for life and serve a wide range of important functions in the body, such as aiding in digestion and metabolism
Enzymes are biological molecules (typically proteins) that significantly speed up the rate of virtually all of the chemical reactions that take place within cells. They are vital for life and serve a wide range of important functions in the body, such as aiding in digestion and metabolism
De novo and salvage pathway of nucleotides synthesis.pptx✨M.A kawish Ⓜ️
This slides explains Metabolism topic "De novo and salvage pathway of nucleotides synthesis. In which synthesis of Purines and pyrimidines synthesis has been occurred. In last there is a difference between these two pathways.
ATP synthase—also called FoF1 ATPase is the universal protein that terminates oxidative phosphorylation by synthesizing ATP from ADP and phosphate.
ATP Synthase is one of the most important enzymes found in the mitochondria of cells
De novo and salvage pathway of nucleotides synthesis.pptx✨M.A kawish Ⓜ️
This slides explains Metabolism topic "De novo and salvage pathway of nucleotides synthesis. In which synthesis of Purines and pyrimidines synthesis has been occurred. In last there is a difference between these two pathways.
ATP synthase—also called FoF1 ATPase is the universal protein that terminates oxidative phosphorylation by synthesizing ATP from ADP and phosphate.
ATP Synthase is one of the most important enzymes found in the mitochondria of cells
Enzyme inhibition is explained with its kinetics, animations showing mechanism of inhibitors action, examples of inhibitors are explained in detail with Enzyme inhibited.
by Dr. N. Sivaranjani, MD
Laboratory method for measuring enzyme activity.
Vital for study of enzyme kinetics and enzyme inhibition.
Measurement of enzyme activity – follow the change in concentration of substrate or product – measure reaction rate.
This power-point presentation will give a complete overview about enzymes, nomenclature of enzymes. Enzymes inhibition is also covered in this ppt. Along with some basin introduction to G- protein coupled receptors is also provided.
Enzymes are biological molecules (proteins) that act as catalysts and help complex reactions occur everywhere in life. Let's say you ate a piece of meat. Proteases would go to work and help break down the peptide bonds between the amino acids.
Enzymes mechanism of action, their specificity types, active center structure and action, inhibitor types, fisher and Koshlend theory are presented. Enzymes classification, a new class of enzymes discovered recently, detailed explanation of each class reaction types is presented as well
These simplified slides by Dr. Sidra Arshad present an overview of the non-respiratory functions of the respiratory tract.
Learning objectives:
1. Enlist the non-respiratory functions of the respiratory tract
2. Briefly explain how these functions are carried out
3. Discuss the significance of dead space
4. Differentiate between minute ventilation and alveolar ventilation
5. Describe the cough and sneeze reflexes
Study Resources:
1. Chapter 39, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 34, Ganong’s Review of Medical Physiology, 26th edition
3. Chapter 17, Human Physiology by Lauralee Sherwood, 9th edition
4. Non-respiratory functions of the lungs https://academic.oup.com/bjaed/article/13/3/98/278874
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Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
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Microteaching is a unique model of practice teaching. It is a viable instrument for the. desired change in the teaching behavior or the behavior potential which, in specified types of real. classroom situations, tends to facilitate the achievement of specified types of objectives.
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
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Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
Explore natural remedies for syphilis treatment in Singapore. Discover alternative therapies, herbal remedies, and lifestyle changes that may complement conventional treatments. Learn about holistic approaches to managing syphilis symptoms and supporting overall health.
Knee anatomy and clinical tests 2024.pdfvimalpl1234
This includes all relevant anatomy and clinical tests compiled from standard textbooks, Campbell,netter etc..It is comprehensive and best suited for orthopaedicians and orthopaedic residents.
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
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40. The Future of Ribozymes In Vitro Molecular Evolution of RNA High Throughput Screening Ribozyme-Based Therapies +
41. In Clinical Trial... HIV Gene Therapy... Bone Marrow Sample Treat Stem Cells with Retroviral Vector Re-Implant Treated Cells Encodes Gene for anti-HIV Ribozyme
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43.
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46. Enzyme Stabilizes Transition State S P ES ES T EP S T Energy change Energy required (no catalysis) Energy decreases (under catalysis) Sub.(S) Prod. (P)Enz(E) T = Transition state V=rate of change of S to P/mt. Adapted from Alberts et al (2002) Molecular Biology of the Cell (4e) p.166 Reaction direction
47.
48. Control Points of Gene Regulation Prokaryotics Post-translational control Eukaryotics Juang RH (2004) BCbasics DNA ribosome mRNA proteins proteins cap 5’ 3’ tail mature mRNA DNA 5’ 3’ process mRNA Translation Activity Proteolysis Transcription RNA Processing RNA Transport RNA Degradation
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50.
51. Active Site Avoids the Influence of Water Preventing the influence of water sustains the formation of stable ionic bonds - +
52. Active Site Is a Deep Buried Pocket Why energy required to reach transition state is lower in the active site? It is a magic pocket (1) Stabilizes transition (2) Expels water (3) Reactive groups (4) Coenzyme helps (2) (3) (4) (1) CoE + - Juang RH (2004) BCbasics
84. Sigmoidal Curve Effect Sigmoidal curve Exaggeration of sigmoidal curve yields a drastic zigzag line that shows the On/Off point clearly Positive effector (ATP) brings sigmoidal curve back to hyperbolic Negative effector (CTP) keeps Consequently, Allosteric enzyme can sense the concentration of the environment and adjust its activity Noncooperative (Hyperbolic) Cooperative (Sigmoidal) v o [Substrate] Off On Juang RH (2004) BCbasics CTP ATP v o
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92. Regulation of Enzyme Activity P R R + proteolysis phosphorylation cAMP or calmodulin or regulator effector (+) Juang RH (2004) BCbasics Regulatory subunit or inhibitor P (-) Inhibitor Proteolysis Phosophorylation Signal transduction Feedback regulation
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103. CCC Allosteric Enzyme ATCase + Active relaxed form Inactive tense form ATCase R R R R R R CCC Catalytic subunits Catalytic subunits Regulatory subunits Aspartate Carbamoyl phosphate Carbamoyl aspartate Juang RH (2004) BCbasics Quaternary structure COO - CH 2 HN-C-COO - H H - - - - O H 2 N-C-O-PO 3 2- = O H 2 N-C- = COO - CH 2 N-C-COO - H H - - - - ATP CTP Nucleic acid metabolism Feedback inhibition CTP CTP CTP CTP CTP CTP
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105. The Reception and Transduction of Signals -GDP +GTP Adenylate cyclase Glycogen Synthase active Insulin kinase Glucagon A The third group: Ion-channel-linked Receptor Gilman, Rodbell (1994) Glycogen breakdown Juang RH (2007) BCbasics SH2 domain G protein GDP + Signal GDP GTP GTP + Signal Activation P Protein Phosphatase Glycogen Synthase P P P P P G-protein-linked Receptor Enzyme-linked Receptor Glycogen
106. Sigmoidal Curve Effect Sigmoidal curve Exaggeration of sigmoidal curve yields a drastic zigzag line that shows the On/Off point clearly Positive effector (ATP) brings sigmoidal curve back to hyperbolic Negative effector (CTP) keeps Consequently, Allosteric enzyme can sense the concentration of the environment and adjust its activity Noncooperative (Hyperbolic) Cooperative (Sigmoidal) v o [Substrate] Off On Juang RH (2004) BCbasics CTP ATP v o
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110. cAMP Controls Activity of Protein Kinase A A A A A cAMP Active kinase CREB CREB Nucleus Activation Gene expression ON DNA Alberts et al (2002) Molecular Biology of the Cell (4e) p. 857, 858 R C R C R R A A A A C C Regulatory subunits Catalytic subunits C P
132. Enzyme Inhibition (Mechanism) Competitive Non-competitive Uncompetitive E E Different site Compete for active site Inhibitor Substrate Cartoon Guide Equation and Description [ I ] binds to free [E] only, and competes with [S]; increasing [S] overcomes Inhibition by [ I ]. [ I ] binds to free [E] or [ES] complex; Increasing [S] can not overcome [ I ] inhibition. [ I ] binds to [ES] complex only, increasing [S] favors the inhibition by [ I ]. X Juang RH (2004) BCbasics E + S -> ES -> E + P + I ↓ E I ← ↑ E + S -> ES -> E + P + + I I ↓ ↓ E I + S ->E I S ← ↑ ↑ E + S -> ES -> E + P + I ↓ E I S ← ↑
134. Enzyme Inhibition (Plots) V max K m K m ’ [S], mM v o I I V max unchanged K m increased V max decreased K m unchanged Both V max & K m decreased I = K m ’ Juang RH (2004) BCbasics K m Competitive Non-competitive Uncompetitive Direct Plots Double Reciprocal V max [S], mM v o K m [S], mM V max I K m ’ V max ’ V max ’ 1/[S] 1/K m 1/ v o 1/ V max I Two parallel lines I Intersect at X axis 1/ v o 1/ V max 1/[S] 1/K m 1/[S] 1/K m 1/ V max 1/ v o Intersect at Y axis
143. Enzyme Inhibitors Are Extensively Used ● Sulfa drug (anti-inflammation) Pseudo substrate competitive inhibitor ● Protease inhibitor Plaques in brain contains protein inhibitor ● HIV protease is critical to life cycle of HIV HIV protease (homodimer): ↑ inhibitor is used to treat AIDS Symmetry Not symmetry -> Human aspartyl protease: (monodimer) Alzheimer's disease domain 1 Asp Asp domain 2 subunit 2 Asp subunit 1 Asp
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150. Intracellular Distribution of Diagnostic Enzymes ACP ALP GGT CK ALP AMS LPS AMS LD 1 AST CK LD 5 ALT AST Prostate Biliary Tract Muscle Bone Salivary Glands Pancreas Heart Liver
182. Enzyme Active Site Is Deeper than Ab Binding Instead, active site on enzyme also recognizes substrate, but actually complementally fits the transition state and stabilized it. Ag binding site on Ab binds to Ag complementally, no further reaction occurs. X
183. cAMP Controls Activity of Protein Kinase A A A A A cAMP Active kinase CREB CREB Nucleus Activation Gene expression ON DNA Alberts et al (2002) Molecular Biology of the Cell (4e) p. 857, 858 R C R C R R A A A A C C Regulatory subunits Catalytic subunits C P
184. HIV protease vs Aspartyl protease Asymmetric monomer ↓ HIV protease (homodimer) HIV Protease inhibitor is used in treating AIDS Symmetric dimer ↑ Aspartyl protease (monomer) Juang RH (2004) BCbasics Asp subunit 2 subunit 1 Asp domain 1 domain 2 Asp Asp
185. Enzyme Inhibitors Are Extensively Used ● Sulfa drug (anti-inflammation) Pseudo substrate competitive inhibitor ● Protease inhibitor Plaques in brain contains protein inhibitor ● HIV protease is critical to life cycle of HIV HIV protease (homodimer): ↑ inhibitor is used to treat AIDS Symmetry Not symmetry -> Human aspartyl protease: (monodimer) Alzheimer's disease domain 1 Asp Asp domain 2 subunit 2 Asp subunit 1 Asp
186. Sulfa Drug Is Competitive Inhibitor Precursor Folic acid Tetrahydro- folic acid Sulfanilamide Sulfa drug (anti-inflammation) Para-aminobenzoic acid (PABA) Bacteria needs PABA for the biosynthesis of folic acid Sulfa drugs has similar structure with PABA, and inhibit bacteria growth. Domagk (1939) -COOH H 2 N- -SONH 2 H 2 N-
187. The Reception and Transduction of Signals -GDP +GTP Adenylate cyclase Glycogen Synthase active Insulin kinase Glucagon A The third group: Ion-channel-linked Receptor Gilman, Rodbell (1994) Glycogen breakdown Juang RH (2007) BCbasics SH2 domain G protein GDP + Signal GDP GTP GTP + Signal Activation P Protein Phosphatase Glycogen Synthase P P P P P G-protein-linked Receptor Enzyme-linked Receptor Glycogen
188. Signal Transduction Network (Ras vs. P53) Cytosol Cell membrane Effector enzyme Signal protein E2F Transcription factor Target gene mRNA Inhibitor P53 Cell division ON Signal Receptor Nucleus Ribosome Transcription Transcription Apoptosis Cell function are controlled by protein interactions mRNA Regulator protein Juang RH (2007) BCbasics Ras
189. The Reception and Transduction of Signals -GDP +GTP Adenylate cyclase Glycogen Synthase active Insulin kinase Glucagon A The third group: Ion-channel-linked Receptor Gilman, Rodbell (1994) Glycogen breakdown Juang RH (2007) BCbasics SH2 domain G protein GDP + Signal GDP GTP GTP + Signal Activation P Protein Phosphatase Glycogen Synthase P P P P P G-protein-linked Receptor Enzyme-linked Receptor Glycogen
190. A PKA active inactive Glucagon P P GP kinase GP kinase GP a GP b Glycogen synthase Glycogen synthase P Protein phosphatase-1 Protein phosphatase-1 Protein phosphatase inhibitor-1 Protein phosphatase inhibitor-1 Glycogen P Phosphatase
191. Classification of Proteases Metal Protease Serine Protease Cysteine Protease Aspartyl Protease Family Example Mechanism Specificity Inhibitor Juang RH (2004) BCbasics Carboxy- peptidase A Chymotrypsin Trypsin Papain Pepsin Renin H57 D102 S195-O - C25-S - H195 D215 D32 H 2 O Non- specific Non- specific Aromatic Basic Non- polar EDTA EGTA DFP TLCK TPCK PCMB Leupeptin Pepstatin E72 H69 Zn 2+ H196
192. Modification of Subtilisin and Its Activity Change No enzyme 1 Asn 155 -> Leu ● ● ● 10,000,000 ( Asn 155 stabilizes transition state ) His & Asp -> Ala ● ○ ○ 37,000 Ser , His & Asp -> Ala ○ ○ ○ 4,000 Subtilisin ● ● ● 10,000,000,000 Active Site Relative Modification Triad: Ser His Asp activity Ser -> Ala ○ ● ● 5,000 Asp -> Ala ● ● ○ 330,000
193. Serine Protease and AchE Chymotrypsin – Gly – Asp – Ser – Gly – Gly – Pro – Leu – Trypsin – Gly – Asp – Ser – Gly – Gly – Pro – Val – Elastase – Gly – Asp – Ser – Gly – Gly – Pro – Leu – Thrombin – Gly – Asp – Ser – Gly – Gly – Pro – Phe – Plasmin – Gly – Asp – Ser – Gly – Gly – Pro – Leu – Acetylcholinesterase – Gly – Glu – Ser – Ala – Gly – Gly – Ala – Chymotrypsin – Val – Thr – Ala – Ala – His – Cys – Gly – Trypsin – Val – Ser – Ala – Gly – His – Cys – Tyr – Elastase – Leu – Thr – Ala – Ala – His – Cys – Ile – Thrombin – Leu – Thr – Ala – Ala – His – Cys – Leu – Plasmin – Leu – Thr – Ala – Ala – His – Cys – Leu – Acetylcholinesterase – – – – – – – – – – – – – His – – – – – – – – Ser 195 Chymotrypsin – Thr – Ile – Asn – Asn – Asp – Ile – Thr – Trypsin – Tyr – Leu – Asn – Asn – Asp – Ile – Met – Elastase – Ser – Lys – Gly – Asn – Asp – Ile – Ala – Thrombin – Asn – Leu – Asp – Arg – Asp – Ile – Ala – Plasmin – Phe – Thr – Arg – Lys – Asp – Ile – Ala – Acetylcholinesterase – – – – – – – – – – – – – – Asp – – – – – – – His 57 Asp 102 Adapted from Dressler & Potter (1991) Discovering Enzymes, p.244
194. H AchE Has Similar Catalytic Mechanism H - O - H H 2 O Adapted from Dressler & Potter (1991) Discovering Enzymes, p.243 ↓ Deacylation Acylation↑ AchE O - C H O CH 3 CH 3 – C – O–CH 2 –CH 2 – N –CH 3 CH 3 + AchE O C H O CH 3 – C CH 3 H O–CH 2 –CH 2 – N –CH 3 CH 3 + AchE O - C H H O CH 3 – C – OH AchE O - C H O CH 3 – C CH 3 O–CH 2 –CH 2 – N –CH 3 H CH 3 +
195. Different Enzymes Might Adopt Same Mechanism Hi, Everybody! ← Useful ↙ Amusing Juang RH (2004) BCbasics O - C Sesame Triad
196. Convergent and Divergent Trypsin Chymotrypsin Elastase Thrombin Plasmin Acetylcholin esterase Thyroglobulin Ester bond Peptide bond hydrolyze acetylcholine Serine Protease Juang RH (2004) BCbasics Divergent evolution Asp-- His-- Ser Asp--His--Ser Convergent evolution C N C C H O C C C O O Evolution Molecular
197. Activity Regulation of Glycogen Phosphorylase Covalent modification P P GP kinase GP phosphatase 1 Non-covalent A A A AMP ATP Glc-6-P Glucose Caffeine Glucose Caffeine spontaneously R T R T Garrett & Grisham (1999) Biochemistry (2e) p.679 P A P A P P A A P A P A P P P A P A
198. CCC Allosteric Enzyme ATCase + Active relaxed form Inactive tense form ATCase R R R R R R CCC Catalytic subunits Catalytic subunits Regulatory subunits Aspartate Carbamoyl phosphate Carbamoyl aspartate Juang RH (2004) BCbasics Quaternary structure COO - CH 2 HN-C-COO - H H - - - - O H 2 N-C-O-PO 3 2- = O H 2 N-C- = COO - CH 2 N-C-COO - H H - - - - ATP CTP Nucleic acid metabolism Feedback inhibition CTP CTP CTP CTP CTP CTP
199. Regulation of Enzyme Activity P R R + proteolysis phosphorylation cAMP or calmodulin or regulator effector (+) Juang RH (2004) BCbasics Regulatory subunit or inhibitor P (-) Inhibitor Proteolysis Phosophorylation Signal transduction Feedback regulation
Acetylcholinesterase (AchE) 是與 Ser protease 家族完全無關的一個酵素,它獨立演化出類似 Ser 催化鐵三角的 Asp-His-Ser 活性區,其作用模式也與 Ser protease 類似,但催化的是水解 acetylcholin 的 ester bond 而非 peptide bond ;事實上,這兩種鍵結的水解方式,也極為相像。這種由不相關的前驅物開始,卻發展出相同的催化機制,稱為 趨同演化 ;而 Ser protease 家族則是由一個相同的始祖開始, 趨異演化 成各個不同的家族成員。 因此,演化現象不但在鉅觀的生物圈發生,同時也在分子層次的微觀世界上演。
肝糖磷解脢 的重要性可由其複雜的調控機制得證,幾乎囊括所有重要的調節方式。因此異位 脢 這種細膩的蛋白質調控方法,當然在肝糖磷解 脢 也有,而且效應物種類繁多。 肝糖磷解 脢 的各種調控機制可以整理成為兩大類,其一為共價性修飾,就是 Ser 14 的磷酸化反應;另一則為非共價性的修飾,正向的活化物只有 AMP 一種,要特別注意並不是 cAMP ;而負面的抑制劑則有許多,例如葡萄糖、 Glc-6-P 、 ATP 與咖啡因。這兩大類效應物真是壁壘分明,例如 AMP 與 ATP 的能量狀況恰好相反,血中的 AMP 濃度高了,表示需要能量,因此活化了肝糖磷解 脢 以便進行磷解反應,取得能量;反之,若血中充滿了 ATP ,則關掉肝糖磷解 脢 活性,不須再降解肝糖。 上圖的右半部需加說明,在磷酸化之後,肝糖磷解 脢 很快由 T 轉換成 R ,因此往下的箭頭較粗。但是即使有磷酸化,當加入負效應物時,也會由 R 轉回 T 而失去活性。
異位脢 的最典型例子,就是 aspartate transcarbamoylase (ATCase) 。此酵素催化上圖的反應,所產生的生成物會繼續代謝,最後生成 CTP 。 此 CTP 會回頭與 ATCase 結合,再迴饋抑制其活性 ( 因為 CTP 太多表示不用再繼續此一代謝路徑了 ) 。因為 CTP 與 ATCase 結合在其 R 次體上,而非 C 次體上的活性區,因此是一種道地的異位 脢 。 CTP 之所以能抑制 ATCase 的活性,是因為當 CTP 結合到 R 次體後,會牽動 C 次體的構形,使得 ATCase 由原來活躍的 relaxed form 轉變成較不具活性的 tense form 。