LONG PRACTICE QUIZ I CANNOT NAME

toxin that inhibits SERCA resulting in reduced ER lumen Ca++

thapsigargin

toxin that inhibits the attachment of the first N-acetylglucosamine during the synthesis of the assembly of the oligosaccharide for N-linked glycosylation

tunicamycin

primary organizing protein associated with all protein-folding activities within the rER lumen

Bip

maintains the monomeric state of IRE and PERK

Bip

exerts a kinase action on eIF2alpha

PERK

expressed protein of the unfolded protein response (aka ER-stress response) most closely associated with linking it to apoptosis

CHOP

anti-apoptotic protein lowered in expression in response to CHOP action

Bcl-2

pro-apoptotic protein raised in expression by p53

Bax

leaks out of the mitochondrion through Bax-ringed "outer membrane permeability pores" with Apaf-1 to form the apoptosome

cytochrome c

dimerized by the apoptosome to cross-cleave and produce two active enzymes

procasapase-9

cuts at the sequence ...DAVD... within ICAD

caspase-3

enzyme directly producing multiple DNA fragments in terminal stages of apoptosis

CAD

stratum \_______ layer of the epidermis whose characteristic appearance (giving rise to its name) results from lowered expression of Bcl-2

granulosum

reactive amino acid within caspase active sites

cysteine

partner amino acid within caspase active sites that withdraws a proton from the reactive amino acid

histidine

used to stabilize the protonated partner during the reaction cycle of caspase-9

glutamate

reporter enzyme used to judge the degree of CHOP regulated expression from a Bcl2-promoter

chloramphenicol acetyl-transferase

assay that most directly monitors the level of transcription of the Bcl2 gene

RT-PCR

used to separate 14-C labeled chloramphenicol from acetylated forms

thin-layer chromatography

live cell membrane-impermeable stain that is not excluded by apoptosed cells

propidium iodide

transgenically expressed clotting factor in CHO cells that induces the unfolded protein response and is poorly secreted

wtFVIII

highly reduced size of the transgenically expressed clotting factor missing the entire fragment which has a large number of targets for N-linked glycosylation

BDD

reduced size of the transgenically expressed clotting factor with only 6 closely spaced targets for N-linked glycosylation within the molecularly modified B domain

226/N6

form of transgenically expressed clotting factor that is best secreted by CHO cells

226/N6

ER lumen protein which bonds to N-linked glycosylations in which terminal glucoses have been retained

calnexin

inhibitor of NOSs which results in higher production of superoxide and less NO

ADMA

modifier of C674 of SERCA that enables glutathiolation

peroxynitrite

modification of SERCA C674 that prevents glutathiolation in atherosclerotic tissue

sulfonation

form of SERCA C674 that is reactive with iodoacetamide

sulfhydryl

modification of C674 that increases SERCA action

gluthiolation

substrate used to identify fragmented DNA with reactive 3'-OHs during TUNEL assays

dUTP-5-fluorescein

enzyme used to label fragmented DNAs in TUNEL assays

terminal deoxynucleotidyl transferase

enzyme that produces reactive 3'-OH's in apoptosis

CAD

virus used to create transformed CHO cell lines and integrate exogenous genes (such as FVIII) for expression

SV40

substance that is fluorescently non-responsive but can be oxidized within the cell by reactive oxygen species to become fluorescently green

dichlorofluorescin-H2

antioxidant found in many food products (notably breakfast cereals) that reduces the unfolded protein response to FVIII overexpression

butylated hydroxyanisole

substance that activates expression from the SV40 late promoter allowing production of transgenic gene products (such as FVIII)

sodium butyrate

used to count the numbers of cells that display colors that are instrumentally detected usually associated with some variable phenomenon

FACS

control applied substance used to oxidize dichlorofluorescein in CHO cells

hydrogen peroxide

protein expressed experimentally in CHO cells that results in the highest DCF fluorescence in CHO cells

wtFVIII

substance used to lower DCF fluorescence in CHO cells caused by overloading the ER

butylated hydroxyanisole

enzyme that reduces hydrogen peroxide using GSH

glutathione peroxidase

oxidized product of glutathione peroxidase

GSSG

breakdown product produced in cells in which peroxide has reacted with lipids

malondialdehyde

SV40 late promoter driven expression of \_______ produces the highest level of ROS

wtFVIII

SV40 late promoter driven expression of which form of a clotting factor produces the lowest level of ROS

226/N6

podocytes (specialized endothelial cells of the glomerulus that sieve out cells and large macromolecules allowign only fluid flow into nephrons) produces what product that reacts with DAF-FM Diacetate to cause it to fluoresce green

NO

ADMA-treated glomeruli take up what provided macromolecule (experimentally) unlike untreated glomeruli

bovine serum albumin

ADMA-treated glomeruli that have taken up provided macromolecule from solution quantifiably swell when now dipped in \____

water

the \____ of a swollen ADMA-treated glomerulus is a measure of its damage

diameter

ADMA damage to glomeruli can be reduced and reversed in response to NO produced by varying concentrations of:

deta-NONOate

NO stimulates what enzyme most directly

guanylate cyclase

what activator of PKG can cross the cell membrane and reduce the degree of ADMA damage to glomeruli

Br-cGMP

what compound oxidizes the heme of soluble guanylate cyclase

ODQ

TEMPOL is a scavenger for \_____ which prevents ADMA damage to glomeruli

superoxide