proteins

transport

besides structural roles, name one other biomedical importance of proteins

hormones

what type of protein acts as a receptor

degradation

proteins undergo life cycle changes including synthesis, maturation, functional regulation, aging, and _________

false

proteins are static molecules with no changes throughout their lifespan

small beads

what is the stationary phase in column chromatography?

increasing ionic strength

how are proteins eluted from ion-exchange chromatography?

Polyacrylamide Gel Electrophoresis (PAGE)

what is the primary method for assessing protein purity?

physiological states or diseases

what can biomaker proteins help diagnose?

size-exclusion chromatography

which chromatography method separates proteins by size/shape?

cation exchanger

what type of ion exchanger binds positively charged proteins?

SDS

what does SDS-PAGE use to denature proteins?

high pressure

HPLC uses tightly packed small beads and requires __________ for separations

excluded from pores

large proteins elute first in size-exclusion chromatography because they are _____________

affinity chromatography

which chromatography separates based on specific ligand binding?

isoelectric point (pI)

what property is used for separation in isoelectric focusing (IEF)?

negatively charged proteins

anion exchangers bind _________________

hydrophobicity

hydrophobic interaction chromatography separates proteins based on ____________

true

affinity chromatography elutes proteins by competition with free ligand or disrution of interactions

false

isoelectric focusing separates proteins based on their molecular weight

Size-Exclusion Chromatography

strokes radius (size/shape)

Ion-Exchange Chromatography

charge-charge interactions

Hydrophobic Interaction Chromatography

Hydrophobicity

Affinity Chromatography based separation principle

specific ligand binding

Sanger

who was the first to determine the sequence of a polypeptide

1-fluoro-2,4-dinitrobenzene (FDNB)

what reagent did Singer use to label N-terminal groups?

sequential reading without destruction

what is the advantage of the Edman Reaction over Sanger’s method?

5-30

what is the practical read length for Edman sequencing per run?

CNBr

what chemical cleavage agent is mentioned for large proteins

DNA sequencing

What method has largely replaced chemical sequencing of peptides?

Amplifies tiny gene amounts

How does recombinant DNA technology aid protein sequencing?

identifies proteins from small sequence data

what is the role of genomics in protein identification?

mass spectrometry

what technique can detect post-translational modifications?

Mass to charge (m/z) ratio

what is the primary separation mechanism in quadrupole analyzers?

Time of flight (TOF)

Which MS analyzer is suited for larger masses like whole proteins?

Tandem Mass Spectrometry

what does MS/MS stand for in tandem mass spectrometry?

Generate sequence-informative fragments

What is the purpose of colliding peptides in MS/MS?

DNA sequencing of gene

a hybrid strategy for protein sequencing involves limited Edman sequencing and the ________________

False

mass spectrometry is less sensitive than Edman chemistry for detecting modifications

True

Electrospray ionization (ESI) is well suited for direct coupling with LC (HPLC)

the full set of proteins in a cell

what does proteomics aim to identify?

with cell type, development, and stimuli

how does the proteome vary?

Post-translational Modifications

What are PTMs in proteomics?

hard to recover enough peptide

What was a limitation of 1st-gen proteomics workflows using SGS-PAGE/2D gels?

Mass spectrometry (MS)

What technqiue is central to 2nd-gen proteomics?

Multi-dimensional chromatography

what does MudPIT stand for in chromatography?

Sensitive analysis of low-abundance peptides

what advantage do moderm MS instruments offer for complex mixtures?

dynamic

the proteome is _________, varying with cell type, development, and stimuli

functional diversity

post-translational modifications (PTMs) mature/regulate proteins and add functional diversity

False

2nd-gen proteomics typically involves isolating individual proteins before mass spectrometry