Aging and Oxidative Damage to Mitochondrial Proteins

Slides:

Advertisements

Similar presentations

- D A N I S H A G I N G R E S E A R C H C E N T E R - Why do we age so differently? Tinna Stevnsner for Christina Poulsen Hvitby (on maternity.

Advertisements

Calmodulin and Phosphorylase Interaction By James Proestos Biochemistry and Biophysics Department Dr. Sonia Anderson’s Lab Agriculture and Life Science.

Mitochondria. Mitochondrial Structure (cont.) Mitochondrial Structure.

The Search For Mitochondrial Ribonucleotide Reductase Daniel Bai Dr. Christopher Mathews Department of Biochemistry and Biophysics HHMI.

Electron Transport and Oxidative Phosphorylation It all reduces down to water.

DNMP Kinase Activity in Mitochondria and Its Role in Mitochondrial Mutagenesis  Brian M. Blair  Dr. Christopher K. Mathews  Department of Biochemistry.

Proteomics of Protein Glutathionylation in Mitochondria of an Aging Heart Jessica Page Mentor: Dr. Claudia Maier Department of Chemistry.

Removal of Unreacted Dinitrophenyl Hydrazine from Carbonyl Derivatives Amira Barkal Mentored by Dr. Gary Merrill Biochemistry and Biophysics.

Analysis of carbonyl- containing compounds derivatized with dinitrophenylhydrazine Thi Nguyen Oregon State University P.I.: Dr. Gary Merrill Biochemistry.

Stage 4: Electron Transport Chain

Oxidative phosphorylation NADH transport Oxidative phosphorylation.

Toxicity of Acetaldehyde with Oxygen Radicals Heather Bolstad Mentor: Joseph S. Beckman, Ph.D. August 28, 2003.

Chapter 3 Energy Transformation Objectives: Review processes and reactions important to metabolic energy transformation Recall the rules that govern energy.

The Disulfide Bonding Pattern of Vaccinia Virus Protein L1R Cliff Gagnier Dr. Dennis Hruby’s Lab September 1, 2004.

Pox Proteomics: Identification of the Proteins Associated with the Membranous Fraction of Vaccinia Virus Cliff Gagnier Dr. Dennis Hruby Department of Microbiology.

Tony Tong Dr. Fred Stevens Dr. Claudia Maier Duane Mooney

OXIDATION PHOSPHORYLATION-2

OXIDATION PHOSPHORYLATION-1 BIOC DR. TISCHLER LECTURE 28.

The Dynein Motor Protein Complex: Structure and Dynamics HHMI Summer Research 2009 with Dr. Elisar Barbar By: Jonathan Yih

Danish Aging Research Center

Oxidative Phosphorylation

Figure 7.UN01 becomes oxidized (loses electron) becomes reduced (gains electron)

Cellular Respiration. CATABOLISM “ENTROPY” ENERGY FOR: ANABOLISMWORK Chemical Potential Energy.

~~ ~~ Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display GLUCOSE Pentose phosphate pathway Starts.

AP Biology Cellular Respiration Part 2. Is Oxygen present?

The Electron Transport Chain & Chemiosmosis. Aerobic Respiration 1.Glycolysis: C 6 H 12 O 6  2C 3 H 4 O ATP + 2 NADH 2.Krebs: 2C 3 H 4 O 3  6CO.

ATP synthesis : The F 1 F 0 -ATPase. Hypotheses on the mechanism of ATP synthesis in mitochondria: “Substrate level phosphorylation” -- coupling of ATP.

Objective: You will be able to compare and contrast the equations of respiration. Do Now: Read p. 221 What is the most important use of food?

Cellular Respiration Part IV: Oxidative Phosphorylation.

Electron Transport Chain and Oxidative Phosphorylation Dr. Sooad Al-Daihan Biochemistry department.

Electron Transport Occurs on the Cristae or intermembrane of Mitochondria.

NADH NAD + H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ O2O2 O H2OH2O H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ H+H+ Electron Transport ATP.

Outer membrane inner membrane matrix red = respiratory chain proteins green = ATP synthases Figure 1. Location of enzymes involved in oxidative phosphorylation.

Chapter 7 Oxidative Phosphorylation. You Must Know How electrons from NADH and FADH 2 are passed to a series of electron acceptors to produce ATP by chemiosmosis.

Stage 4: Electron Transport Chain (ETC) and Chemiosmosis

AP Biology  convert glucose (6C) to 2 pyruvate (3C)  produces: 4 ATP & 2 NADH  consumes: 2 ATP  net: 2 ATP & 2 NADH glucose C-C-C-C-C-C P-C-C-C-C-C-C-P.

CHAPTER 7: CELLULAR RESPIRATION. CELLULAR RESPIRATION Process where cells make ATP by breaking down glucose.

Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Cellular Respiration: Harvesting Chemical Energy.

PJK Using HX-MS to Determine Protein Structure of AhpC 2, a Peroxiredoxin Piper J. Klemm Faculty Advisor: Claudia S. Maier, PhD HHMI Fellowship,

LEHNINGER PRINCIPLES OF BIOCHEMISTRY

The Electron Transport Chain

AP Biology Cellular Respiration Overview Part 1. Process of Cellular Respiration.

Citric acid cycle and Oxidative phosphorylation Student.

An Overview of Cellular Respiration

AP Biology Cellular Respiration – Glycolysis, Krebs Cycle, and ETC Part 2.

Dinitrophenyl Hydrazine from Carbonyl Derivatives

Electron Transport & Oxidative Phosphorylation

23.2 Electron Transport and ATP

Electron Transport Chain Manipulative

Cellular Respiration.

Cellular Respiration Stages 2-4.

Chapter 10 Chem 341 Suroviec Fall 2016.

Cellular Respiration Part IV: Oxidative Phosphorylation

Ch 9 (Part 3): E.T.C./ Oxidative Phosphorylation

The Actions of the Three Proton Pumps and ATP Synthase

It’s a big bright beautiful world

Cellular Respiration.

Cellular Respiration Chapter 6.

Mammalian versus yeast OXPHOS system.

Cellular Respiration Part 2

5.7 Electron Transport Chain

Mitochondrial Matrix Reloaded with RNA

Cellular Respiration.

Cellular Respiration Part 2

Cellular Respiration.

It all reduces down to water.

Volume 7, Issue 8, Pages (August 2000)

Mitochondria: important target for drug toxicity?

Figure 1 Oxidative phosphorylation

Chapter 7: Cellular Respiration

Presentation transcript:

Aging and Oxidative Damage to Mitochondrial Proteins Tony Tong Dr. Claudia Maier Dr. Fred Stevens Department of Chemistry Department of Pharmaceutical Sciences Department of Biochemistry and Biophysics Oregon State University August 26, 2005 HHMI Summer 2005 Fellowship Program

Oxygen radicals and aging ROS (H2O2, ·O2-, ·OH) Oxidative damage Heart disease and aging Source: http://onlinetc.its.brooklyn.cuny.edu/ Core81/chap3.html Overall goal: Characterize oxidative damage to mitochondrial proteins

Oxidative Phosphorylation Cell cytosol Outer membrane Intermembrane space Inner membrane Matrix Oxidative Phosphorylation H+ H+ H+ e- III I IV V O2 + H+ e- 4 + 4 2 H2O ADP + Pi H2O2, ·O2-, •OH ATP NADH Adapted from Cooper, G. and Hausman, R., Cell: A Molecular Approach, 2003

Formation of LPO products Linoleic acid ROS (H2O2, ·O2-, ·OH) Aldehyde functionality group 4-hydroxy-2-nonenal (4-HNE) 4-oxo-2-nonenal (4-ONE)

Oxylipid adduction to proteins Cysteine electrophilic C-3 Histidine 4-HNE Lysine

Oxidative Phosphorylation Cell cytosol Outer membrane Intermembrane space Inner membrane Matrix Oxidative Phosphorylation H+ H+ H+ e- III I IV V O2 + H+ e- 4 + 4 2 H2O ADP + Pi H2O2, ·O2-, •OH ATP NADH Adapted from Cooper, G. and Hausman, R., Cell: A Molecular Approach, 2003

Probing for damaged proteins ARP (Aldehyde-reactive probe) hydroxylamine biotin 4-HNE-adducted protein Aldehyde functionality group

Thioredoxin as an in vitro model

Thioredoxin as an in vitro model: Experimental Approach + HNE & ARP Thioredoxin + Trypsin Thioredoxin peptides

MS of ARP-labeled thioredoxin peptides 734.4 HNE-ARP [IIHLTDDSFDTDVLK]3+ + HNE 630.3 HNE [IIHLTDDSFDTDVLK]3+ 577.9 [IIHLTDDSFDTDVLK]3+ m/z

MS/MS of thioredoxin peptides I-I-H-L-T-D-D-S-F-D-T-D-V-L-K b1 b2 b3 b4 b5 b6 b7 b8 b9 b10 b11 b12 b13 b14 y14 y13 y12 y11 y10 y9 y8 y7 y6 y5 y4 y3 y2 y1 N-terminus C-terminus 200 400 600 800 1000 1200 1400 1600 1800 100 % m/z I-I HNE-ARP H L T D S F T-D V b2 227.1 b3 833.5 b7 1278 b6 1163 b8 1365 b9 1512 b10 1627 b12 1843 b13 1942 b4 946.6 b5 1048

Affinity chromatography = ARP ARP (Aldehyde-reactive probe) hydroxylamine biotin Avidin bead column

Affinity chromatography (cont’d) = ARP Avidin bead column

MS of unlabeled peptides 100 MIAPILDEIADEYQGK 1805.9 % 799.0 1441.8 2084.6 2727.4 3370.2 4013.0 m/z

MS of labeled peptides IIHLTDDSFDTDVLK SDKIIHLTDDSFDTDVLK 2201.1 100 IIHLTDDSFDTDVLK HNE-ARP % SDKIIHLTDDSFDTDVLK HNE-ARP 2201.1 2531.3 799.0 1441.8 2084.6 2727.4 3370.2 4013.0 m/z

Future work Maximize recovery of material from beads Repeat with labeled mitochondrial samples Find which and where proteins have been adducted

Acknowledgements The labs of: Dr. Claudia Maier Dr. Fred Stevens Dr. Mike Schimerlik Dr. Emily Ho EHSC Mass Spectrometry Core Facility Dr. Kevin Ahern Dr. Chris Mathews Howard Hughes Medical Institute Sacrificed rats

Questions?

Avidin-immobilized beads acrylamide monomeric avidin azlactone linker

Affinity chromatography ARP-labeled Complex IV ARP LPO IV avidin biotin reversible bond avidin biotin irreversible bond