Sections
Protein Purification
Protein Extraction
Differential Centrifugation
Salting Out
Dialysis
Column Chromatography
Ion-Exchange Chromatography
Anion-Exchange Chromatography
Size Exclusion Chromatography
Affinity Chromatography
Specific Activity
HPLC
Spectrophotometry
Native Gel Electrophoresis
SDS-PAGE
SDS-PAGE Strategies
Isoelectric Focusing
2D-Electrophoresis
Diagonal Electrophoresis
Mass Spectrometry
Mass Spectrum
Tandem Mass Spectrometry
Peptide Mass Fingerprinting
Overview of Direct Protein Sequencing
Amino Acid Hydrolysis
FDNB
Chemical Cleavage of Bonds
Peptidases
Edman Degradation
Edman Degradation Sequenator and Sequencing Data Analysis
Edman Degradation Reaction Efficiency
Ordering Cleaved Fragments
Strategy for Ordering Cleaved Fragments
Indirect Protein Sequencing Via Geneomic Analyses

Concept #1: Overview Of Direct Protein Sequencing

Practice: Which of the following is a protein sequencing technique?

Concept #2: Overview of Techniques for Protein Sequencing

Practice: Appropriately match each option to each description. Options may be used more than once.

a. Cyanogen bromide (CNBr).              _____ An enzyme that cleaves specific peptide bonds.

b. Edman Degradation.                        _____ Nonspecifically cleaves all peptide bonds.          

c. Elastase.                                           _____ A chemical that breaks specific peptide bonds.

d. 6M HCl.                                             _____ Used as an N-terminal protein sequencing technique.

                                                              _____ Used as a peptide cleavage technique.

                                                              _____ Results in smaller peptide fragments.

                                                              _____ Results in free amino acids.

Practice: Appropriately match each option to each reagent. Options may be used more than once.

a. Amino acid hydrolysis.                           _____ Hydrazine.

b. Chemical cleavage.                                _____ Trypsin.           

c. Peptidase.                                              _____ 6M Hydrochloric acid.

                                                                   ______ β-mercaptoethanol + iodoacetate.

                                                                   ______ Chymotrypsin.