1. electrostatic attraction of charged residues (salt bridges/repulsive effects)
2. internal H-bonding (specific distance and angle requirements)
3. short range Van der Waals interactions and London dispersion forces of neutral charged residues
4. entropy driven hydrophobicity (Gibbs free E change)

aspartic acid, glutamic acid, tyrosine, cysteine have negative charges

lysine, arginine, and histidine have positive charges

Zinc fingers

loop of 23 AA resulting from Zn ion being complexed to 4 AA, usually cysteines and histidines

Kringle domains

conserves seq that fold into large loops that are stabilized by 3 disulfide linkages. Impt in protein-protein interactions with blood coag factors.

Leucine Zippers

arrangments of leucines along one side of an alpha-helix btwn 2 proteins, so that protein can form dimers leaving basic AA regions to bind DNA

Ways Proteins Can Be Denatured

1. detergents = straightens out protein (Na dodecyl sulfate makes peptide rodlike
2. chaotropic agents - remove water to unsolubilize protein
3. high temps (50-60 degrees C or higher)
4. drastic pH changes
5. organic solvents

right shift caused by:

decreased pH (increased [H+])

increased [CO2]

increased [2,3-BPG]

increased temperature

all factors lead to increased O2 unloading which means that a higher percentage of O2 is delivered to tissues

Deoxy Hb is usually in taut (T) form where heme groups restricted and hard for O2 to bind

When O2 binds to 1st heme group, the Fe of that heme will shift and pull attached his along.

Movement breaks salt bridges pushing Hb into relaxed (R) state and allow more O2 binding sites to become available

A right shift when pH decreases (increase in H+ conc.)

O2 release from Hb will increase when pH is lowered or [CO2] is high

Hb has decreased O2 affinity

Raising pH or decreasing [CO2]-shift curve to left

Mb has a greater affinity at lower [O2] than Hb

Hb can transport more molecules of O2 b/c of cooperative effect of multiple binding sites

Hb can't readily bind 1st O2, but once it does has higher affinity for O2 at other sites

Hb-sigmoidal curve

Mb-hyperbolic

Hb responds more readily to small changes in partial pressures of O2

Mb saturated quicker than Hb at low partial pressure of O2 (doesn't have much flexibility)

weaken strength of heme-O2 interaction. Heme binds O2 so strongly it can convert it to a superoxide anion, which can be an unwanted oxidizing agent

prevents interaction of oxy-heme groups with other oxy-heme groups

2,3-BPG binds to deoxy Hb-makes it more stable and resistant to oxygenation after O2 delivery

negative charge allows BPG to bind to a site between two alpha chains

causes right shift in O2-dissociation curve (increase O2 release)

Fetal Hb has two gamma chains instead of beta chains.

2,3-BPG can't bind to fetal Hb and it has greater affinity fo O2 and takes O2 from maternal Hb

HbF is left shifted on O2-dissociation curve compared to maternal Hb

Hb binds NO at heme Fe and cys residues in globin chain.

NO relaxant for vascular smooth muscle

Hb can pick up NO, stiffen vascular tissues, and increase BP

NO is strong vasodilator so Hb can be used to mediate vascular tone

crystallization and stiffening of Hbs causes them to adhere to the endothelium and often obstruct small vessels (capillaries) or cause hemolysis

hemolysis can cause anemia and hyperbilirubinemia, causing pallor and jaundice

Complete vasocclusion can cause ishemia or infarction resulting in organ damage

Stroke, infection and excrutiating pain can result

a protein catalyst for specific biochem rxns.

It will not undergo chem rxn itself and can be reused for other rxns

What does enzyme effect and not effect in rxn?

Enzymes reduce activation energy.

Keq and delta G are not affected by enzymes.

lock and key model

describes an enzyme active sites (lock) being specific for binding substrate (key)

describe specificity

induced fit model

active site will conform to fit substrate

describes specificity and catalytic activity

non-protein organic molecule that is tightly bound to enzyme active site

ex-heme, biotin, or flavin

holoenzyme-prosthetic group attached

apoenzyme-prosthetic group NOT attached

oxidoreductases-catalyze RED/OX reactions

Ex-H+ donor to substrate

transferases-transfer group from one molecule to another

Ex-phosphorylases

hydrolases-hydrolytic cleavage of bond

Ex-peptide bond cleavage

lyases-cleave bonds w/o water to leave double bond or addition of other groups to double bond

Ex-decarboxylase

isomerases-change geometry of molecule

Ex-cis-trans isomerase

ligases-join 2 molecules together though hydrolysis of high energy bond

Ex-carboxylase

V = V_max[S]/(K_m+[S])

V - velocity of rxn

Vmax - max rxn velocity

[S] - substrate concentration

Km - constant value for a specific substrate

Vmax - when [S] is very high

Km - [S] when V = 1/2(Vmax)

The LB graph plots the inverse of the Michaelis-Menten plot, which is 1/V on the y-axis and 1/[S] on the x-axis

x-intercept - 1/Km

y-intercept - 1/Vmax

bind directly to the active site and can be overcome by increased [substrate].

Other inhibitors bind to other sites on enzyme

same y-intercept

different x-intercepts

different y-intercepts

same x-intercepts

different y-intercepts

different x-intercepts

bind with ES complex.

irreversible inhibitors.

amount of tissue producing enzyme

rate of enzyme released

rate of enzyme inactivation/elimination from plasma

Aspartate aminotransferase (AST) - catalyzes exhange of amino group between alpha amino acids (aspartate) and alpha keto acids

Alanine aminotransferase (ALT) - catalyzes exchange of amino group between alpha amino acids (alanine) and alpha keto acids

AST - liver damage and myocardial cells,hemolysed blood

ALT - liver damage

Glutathione + amino acids → γ-glutamyl-amino acid + cys-gly

γ-glutamyltranspeptidase found in kidney, intestine, liver, and fetal tissue

GGT levels are elevated and very sensitive in liver damage

Alkaline phosphatase (ALP)

Hydrolyzes phosphate groups from organic monophosphate at pH~9

Found in liver, bone, intestine, and placental tissue

Elevation of ALP means damage to liver and bone tissue

Elevated in gowing children, bone fractures, and pregnant women

Acid Phosphatase

hydrolyzes phosphate groups from organic monophosphates at pH~5

found in prostate, platelets, and erythrocytes

Elevation found in metastatic prostate cancer and hemolysed blood samples

Most common form is LDH2 (HHHM)

After MI or hemolysis, LDH1 (HHHH) predominates causing a "flipped" LDH b/c LDH1>LDH2

Lactate + NAD → pyruvate + NADH + H

interconversion of pyruvate and lactate

Another name for alpha hydroxybutyrate dehydrogenase (HBD)?

has 2 subunits, M and B

MB-MI or brief ischemia

MM-vigorous exercise, injection, convulsions

BB-neural tissue damage

pancreatitis

morphine

starch + H2O → maltose, maltotriose & limit dextrins

digest starch

pancreatic pathology

morphine

trigylceride + H2O → fatty acids + glycerol

fat digestion

proteins that function in the contractile apparatus of the muscle

Cardiac troponin I is increased in plasma 3-6 hours after MI

mildly elevated alkaline phosphatase (healing bone)

elevated creatine kinase MM (skeletal muscle)

mildly elevated alkaline phosphatase (bone growth)

elevated GGT

integrins

immunoglobulin superfamily

cadherins

selectins

NCAM - neural cell adhesion molecule

PECAM - platelet-endothelial cell adhesion molecule

VCAM - vascular cell adhesion molecule

Named for their similarity in structure to the Igs

Rheumatoid arthritis - overexpression of VCAMs

Psoriasis - form of an integrin

Pemphigus vulgaris - autoantibodies interacting with desmoglein

von Willebrand disease - expression of P-selectin

regions of same chain or neighboring chains bond to each other w/ H bonds, the H bonds are perependicular to the long axis of the chain

most stable conformation - antiparallel sheets (one side N→C other side C→N

H bonding of AA 3 sequences apart in same chain

results in U turn bend of chain

ser

thr

tyr

acetylation of N terminus

carboxylation - add COOH

hydroylation - add OH

glycosylation - add glucose

phosphorylation - add phosphate group

disulfide linkages

alpha helix

beta-pleated sheets

beta-turns

random coiling

results from free roation of bonds besides peptide bonds

secondary structures fold on each other

hydrophobic - inside

hydrophilic - outside

fibrous and insoluble

globular and soluble

firbrous and soluble

structural roles

enzymes

role in contractile structures

transport (Hb)

hormones

receptors

Abs

peptides - moderate chain length polymers of amino acids joined by peptide bonds

proteins - polymeric compounds composed of AA joined by peptide bonds

due to presence or absence of N-acetyl-galactosamine or galactose linked to penultimate galactose by C1-C3 linkage

1 sugar difference

unbranched polysacc w/ repeating disacc - one is AA, other is uronic acid

Ex - hyaluronic acid and heparin

maltose - alpha linked

sucrose - alpha linked

lactose - beta linked

starch - alpha linked (spiral)

glycogen - 2 types - alpha C1-C4 (linear)

                                 alpha C1-C6 (branched)

cellulose - beta linked (linear)

alpha and beta linkage affects the shape of the compound and humans can't digest cellulose b/c beta linkages

anomeric -OH group of sugar reacts w/ another -OH compound

amino sugar - -OH group replaced by amino group

Ex - glucosamine, galactosamine

deoxy sugar - 1 OH group replaced by H

Ex - 2-deoxyribose

configuration changes in molecule by ring opening and closing

occurs at anomeric C

pyranose - 6 C ring

furanose - 5 C ring

glucose - most impt monosacc

Polysacc made up of glucose - starch

                                             cellulose

                                             glycogen

fingerprints are different of two twins - different positions in womb

type I diabetes - if one gets, less than 1/2 the time other twin gets

What percentage of live births suffer from a disease causing monogenic defect?

normal female - 2 X chromosomes

normal male - 1 X chromosome

normal female - 0 Y chromosomes

normal male - 1 Y chromosome

trisomy 13

trisomy 18

trisomy 21

trisomy X

XYY

trisomy XYY

trisomy XXX

tall

other mild symptoms

male

testicular failure

can't conceive

X-

sexually immature

short

web neck

phenotypically female

lack of male to male transmission

gender-dependent

x-linked gene comes from mother on X chromosome only, but can affect both males and females, through mostly males show disease

autosomal come from chromosomes other than sex chromosome

An autosomal disease passes on one allele from the mother and one allele from the father

A mitochrondrial disease has only one allele from the female/mother only and all offspring will show some degree of the disease

Which will probably have greatest number of offspring who suffer from an inherited disease (may be more than one):

A. parents both heterozygous for autosomal recessive disease

B. parents both heterozygous for an autosomal dominant disease

C. a father who suffers from an X-linked recessive disease and a mother who is homozygous wild type (has only well type of disease gene)

D. a mother who is heterozygous for an x-linked recessive disease and a father who is wild type hemizygous (has the well form of that disease gene)

E. a father who suffers from a mitochondrial disease and a mother who does not at all have it

Of these, which will probably have the least number of offspring who suffer from an inherited disease (may be more than one):

A. parents both heterozygous for autosomal recessive disease

B. parents both heterozygous for an autosomal dominant disease

C. a father who suffers from an X-linked recessive disease and a mother who is homozygous wild type (has only well type of disease gene)

D. a mother who is heterozygous for an x-linked recessive disease and a father who is wild type hemizygous (has the well form of that disease gene)

E. a father who suffers from a mitochondrial disease and a mother who does not have it

E. a male who suffers from mitochondrial disease and a female who does not

C. a father who suffers from an x-linked recessive disease and a mother who is homozygous wild type (has only well type of disease gene)

Genotype?

Phenotype?

genotype-genetic makeup

phenotype-physical manifestation

used to be thought rare

now known as the most common hereditary disease in the USA

gender - females under 50 not have disease (menstrual cycle expels excess iron)

blood donations

diet - not enough Fe normally and you make most of Fe intake

alcohol consumption

environment

X-inactivation and that males with the defect do not reproduce.

Females need both x's to be defective in order the x-recessive diseases to show

females have two x's

a triplet repeat of CAG (glutamines) amino acids that encode for polyglutamine tract.

Normally - ≤ 34

Have Disease - ≥ 37

lung infections caused by P. aeroginosa

accumulates in lungs and destroys lining

CFTR used to clear out bacteria from lungs

dysfunctional in CF and can't remove bacteria anymore

lungs get destoyed by inhaled bacteria (P. aeroginosa)

cystic fibrosis - protect from typhoid fever

sickle cell anemia - protects from malaria

mental retardation

blindness

paralysis

muscle atrophy

cherry red spot on retina

hypertension

cardiovascular disease

type 2 diabetes

anencephaly

spina bifida

cleft lip/palate

alchoholism

asthma

bipolar disorder

inherited epilepsy

idiopathic gout

obesity

schizophrenizia

type I diabetes

glucocorticoid-remediable aldosteronism

apparent mineralocorticoid excess

Liddle syndrome

PPAR-gamma

glucokinase

calpain 10

Which of the following infectious diseases is NOT thought to be protected against by an inherited genetic mutation or deletion (may be more than one):

A. HIV

B. typhoid fever

C. anthrax

D. malaria

E. leprosy

anthrax

leprosy

single nucleotide polymorphisms

1:300 bps

A patient has the form of the gene that cause Huntington's (20 repeats). The patient asks about a "gray area" in testing and whether the result in certain.

What do you say?

You are treating a case of erythroblastosis fetalis, a disease of the fetus in pregnancy. This is most often caused by isoimmune reaction to the D antigen, which is one of the antigens responsible for the Rh blood group. Abs developed by the Rh- mother "attack" an Rh+ fetus's RBCs leading to hemolysis. You want to quickly counsel the expecting couple about the likelihood the offspring will have another Rh+ fetus. The antigen is inherited in autosomal domianant fashion so you know the mother is

homozygous for the genes that make her Rh-. The father knows he is Rh+ so you able to advise them that on avg?

AND

You ask the father if he has previously fathered an Rh- child and he says YES you are able to advise that on avg?

AND

if the father says that has  has fathered an Rh- child and he instead says NO you are able to advise that on avg?

the info given you can't determine the likelihood of the disease for their future offspring

the info given you can't determine the likelihood of the disease for their future offspring

50% of their offspring will suffer from the disease

autosomal recessive homozygote

male and female

which get disease?

autosomal recessive heterozygote

male and female

which get disease?

autosomal dominant homozygote

male and female

which get disease?

autosomal dominant heterozygote

male and female

which get disease?

male

x-linked recessive heterozygote

male and female

which get disease?

x-linked recessive homozygote

male and female

which get disease?

Mitochondrial, only father has disease

male and female

which get disease?

Mitochondrial, only mother has disease

male and female

which get disease?

blood

saliva

cheek swabs

chorionic villus sampling (CVS) - 0.5-1.0% loss

ultrasound - 0% loss

cordocentesis- 1-2% loss

How many inherited diseases does FL screen for?

What do these diseases have in common that makes them a good idea to screen for?

What instrument made possible the recent expansion in the number of diseases screened in FL and does it analyze sequences or metabolites?

35 diseases screened in FL

cause early damage and effective txt exists

tandem mass spectrometry-analyzes metabolites

may catch a patient gives a false - by another method

easier, less expensive to do in quantity than biochem assays

heritage and fam history of person gives info about what diseases and mutations to look for

it is predictive (before symptom onset)

false negatives due to gene mutation being in unexpected location in gene

limited coverage of test - only test for specific disease, might miss others

may have added expenses due to patented genes

% frequency of two most often occuring mutations in CTFR genes

Any high frequency mutations in familial hypercholesterimia genes?

70% and 2.5%

no, high freq mutations in FH genes

Knowledge of person's VKORC1 and CYP2C9 seq can help you decide what about a patient?

What other seq can be screened?

drug dosage and type

P450 screened also

bacteria

viruses

inherited predispositions

gene defects - always involved

enviro causes

hMLH1

hMSH2

protooncogene b/c it is a growth-stimulating gene

can become oncogene if its activity starts to result in uncontrolled growth

Gene A loses activity w/ certain mutation.

Is the normal gene a tumor suppressor, oncogne, anti-oncogne, or proto-oncogene?

protein/enzymes that require a metal (Zn or Ca) ion to break down proteins in ECM of cell

involved in metastasis

only some cells in tumor can generate new tumor

if can treat or cut out those cells, then localize tumor and control metastasis

[ligand] for 1/2 max occupancy of receptors

measure affinity of ligand for receptor

[ligand] for 1/2 max response

effectiveness in eliciting cellular response

some reside in cytoplasm until encounter ligands-glucocorticoid and aldosterone-Rs

most reside full time in nucleus

receptor ligand complex acts on nucleus by binds to DNA and affecting expression (turn on/off expression)

fatty acid transport

bilirubin transport

transport of steroid hormones

transport of sulta drugs, penicillin, aspirin

most abundant alpha-1 globulin?

Function?

Clinical result of its absence?

alpha-1 antiprotease

protease inhibitor on compounds such as elastase and collagenase

prevents proteolysis in lungs

lung loses ability to recoil after inspiration leading to emphysema and respiratory failure

cortisol-binding globulin (alpha-1 globulin)

sex hormone binding globulin (SHBG)

beta-globulin

iron would be lost due to Hb breakdown.

Hb is lost through the kidney and excreted in urine.

ceruloplasmin

blue b/c of copper

oxidize Fe 2+ Fe 3+ in plasma

low levels found in Wilson's disease