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Biol 432 > Peptide hormones: mechanisms & biosynthesis > Flashcards

Peptide hormones: mechanisms & biosynthesis Flashcards

1
Q

Protein/peptide hormones =

A

most numerous type of hormone

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2
Q

• Many peptide hormones belong to ….

A

families that share genetic and
peptide structure homologies

  • Evolutionary emergence from a common ancestral gene
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3
Q

Most peptide/protein hormones require the

__ of a __ gene

A

Most peptide/protein hormones require the
transcription of a single gene
• e.g., human INS gene (insulin)

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4
Q

• Most peptide/protein hormones require the
transcription of a single gene
however exceptions include…

A

Glycoprotein hormones

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5
Q

Glycoprotein hormones examples (4)

A

• Follicle-stimulating hormone (FSH), Luteinizing
hormone (LH), Thyroid-stimulating hormone
(TSH) and human chorionic gonadotropin (hCG)

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6
Q

Glycoprotein hormones are Heterodimers …..

A

-Heterodimers consisting of alpha and beta
subunits (encoded by two different genes),
with carbohydrate side-chains
• Alpha subunit: same for all four of these
hormones; Beta subunit: unique for each
hormone (and confers biological specificity)

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7
Q

Genes: ____ regions (exons + introns) and ___ regions

A

• Genes: coding regions (exons + introns) and regulatory regions

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8
Q

promoter region

A

• Most important regulatory region: promoter region on 5’ end of gene… includes an
initiator element (w/ transcription start site, generally just downstream of TATA box)
and regulatory elements (specific binding sites for transcription factors: proteins that
may initiate, activate, or repress transcription)

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9
Q

transcription factors

A

transcription factors: proteins that

may initiate, activate, or repress transcription)

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10
Q

what is the most important regulatory region

A

Most important regulatory region: promoter region on 5’ end of gene

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11
Q

wha are enhancers/silencers

A

Enhancers/silencers (regulatory regions that are binding sites for activators or
repressors) are usually located further upstream or downstream of the promoter

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12
Q

Some hormones cause activators or repressors to bind to regulatory elements,
___ or ____ transcription of target genes as part of their cellular effects.

give an example

A

Some hormones cause activators or repressors to bind to regulatory elements,
increasing or decreasing transcription of target genes as part of their cellular effects.
e.g., estrogen receptor binds to specific DNA sequences: estrogen response elements
(EREs). Hormones can both affect and be affected by regulation of gene expression.

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13
Q

what is DNA

A

DNA: tightly packaged (e.g., histone proteins forming nucleosomes, which bundle together)

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14
Q

what is chromatin remodeling

A

• To allow dynamic access to condensed DNA, chromatin remodeling alters this architecture to
expose or hide regions for transcriptional regulation.

Chromatin modeling is important for controling dna expression

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15
Q

chromatin remodeling could involve

A

Histone modifying enzymes,
ATP-dependent chromatin
remodeling complexes

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16
Q

Histone modifying enzymes example

A 
Histone modifying enzymes,
e.g., histone acetyltransferases
(HATs) add acetyl groups to
histones, causing more open
conforma=on (vs. histone
deacetylases, HDACs)
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17
Q

ATP-dependent chromatin

remodeling complexes

A 
• ATP-dependent chromatin
remodeling complexes which
can move, eject or restructure
nucleosomes to expose
binding sites for transcription
activators or repressors at
gene promoters or enhancers

ATP-dependent chromatin remodeling complexes can be
recruited to specific sites such as gene promoters, and
cooperate with histone modifying enzymes.

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18
Q

_____ located far from the

transcriptional start site can impact gene expression

A

Distal regulatory elements located far from the
transcriptional start site can impact gene expression
• They can come into closer proximity due to
changes in the 3-D structure of DNA & chromatin

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19
Q

In the promoter region, the

preinitiation complex forms:

A 
Incl. RNA polymerase II (the
enzyme that transcribes
genes to mRNA), general
transcription factors, and
additional regulators (e.g.,
specific transcription factors,
chromatin remodeling
complexes, etc.)
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20
Q

what influences the rate of gene transcription

A

Presence of regulators & interactions between regulators and

DNA regulatory elements influence the rate of gene transcription

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21
Q 
Once transcription is initiated,
\_\_\_\_ continues the
elongation process until a
sequence is reached and
transcribed that signals \_\_\_\_
and \_\_\_\_\_ of the RNA,
leading to \_\_\_\_\_
A 
Once transcription is initiated,
RNA polymerase II continues the
elongation process until a
sequence is reached and
transcribed that signals cleavage
and polyadenylation of the RNA,
leading to transcription
termination
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22
Q

Pre-mRNA/mRNA processing:

A 
(during transcription): 5’ end
of RNA is capped with modified
guanine nucleotide
(7-methylguanosine)
• Key for nuclear export,
translation and stability
• 3’ end is polyadenylated
(adenosine residues added)
• Key for nuclear export,
translation and stability
• Introns (intervening sequences),
spliced out, and exons
(expressed) incl. 5’ and 3’ UTRs
(untranslated regions) ligated
• Mature mRNA is exported via
nuclear pore
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23
Q

Protein and mRNA are degraded at ___ rates
(often fast for signalling molecules like hormones)
_____ and
___ of 3’ poly-A tail
affect ____ half-life

A

Protein and mRNA are degraded at variable rates
(often fast for signalling molecules like hormones)
5’ mRNA cap and
length of 3’ poly-A tail
affect mRNA half-life

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24
Q

Calcitonin gene-related peptide

A

37-a.a. peptide primarily
secreted in nervous system.
Vasodilator, implicated in
sensing pain

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25
Q

Calcitonin

A

32-a.a. peptide hormone
secreted by the thyroid gland. Reduces
blood calcium, opposing the effects of
parathyroid hormone.

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26
Q

Alternative splicing regulates___

A

Alternative splicing regulates the synthesis of

(typically tissue-specific) hormone or receptor variants

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27
Q 
Hormone \_\_\_ (as well as specific blood binding proteins) are
comprised of proteins, therefore \_\_\_ by genes…
A

Subject to the same types of regulatory control of their gene’s
transcription/translation as peptide hormones
• Some hormones affect the synthesis of their own receptors
(feedback regulation)
•

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28
Q

Hormones might regulate their target genes at ___,

___ and____ levels

A

Hormones might regulate their target genes at transcriptional,
posttranscriptional and posttranslational levels

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29
Q

initiation usually involves….

A 
initiation usually involves
interactions of certain key
proteins (the iniHaHon
factors) with the 5’ mRNA
cap. These proteins bind the
small (40S) ribosomal subunit
and hold the mRNA in place
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30
Q

Control point

A

Control point: forma4on
of ini4a4on complex
during translation

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31
Q

transcription

A

• The mature mRNA is used as a template for the assembly of
amino acids via tRNAs
• Translation of mRNA to protein starts at a specific site on
mRNA = start codon (usually AUG, encodes methionine)
• Translation will continue until a stop codon is reached

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32
Q

Initiation stage

A

: ribosome assembles around the mRNA, and

the first tRNA is attached at the start codon

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33
Q

Elongation stage:

A

peptide bond between methionine and
subsequent encoded amino acid, and the ribosome then
translocates three nucleotides along the mRNA

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34
Q

Termination stage

A

when a stop codon is reached, the

ribosome releases the polypeptide

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35
Q

Translation

A

= very energetically expensive, and is driven by

GTP and ATP hydrolysis

36
Q

____ hormones are stored in &
secreted from ___ – therefore,
must be synthesized within the ____

A

Protein/peptide hormones are stored in &
secreted from secretory vesicles – therefore,
must be synthesized within the rough ER

37
Q

First amino acids that are translated from the

mRNA template form a ___

A

First amino acids that are translated from the

mRNA template form a signal sequence

38
Q

signal sequence causes ….

A

• This causes the nascent peptide chain to enter
a secretory pathway at the beginning of its
formation. The signal sequence bind to a signal recognition particle (SRP), which docks with an SRP receptor complex. A translocation channel transports the nascent peptide chain into the ER lumen; translation continues)

39
Q

Most peptide hormones: ….

A

Most peptide hormones: synthesized as large precursors, which then undergo several
modifications during and/or after translation
• e.g., preprohormone (w/ signal sequence) → prohormone → mature hormone

40
Q

After the transfer of ribosomes to the ____, the growing polypeptide chain enters the ____ as translation continues, and the ____ is rapidly cleaved enzymatically

A 
• After the transfer of ribosomes to the ER membrane, the growing polypeptide chain enters the ER lumen as translation continues, and the signal sequence is rapidly cleaved enzymatically
(in some cases, the
polypeptide generated is
the mature hormone; in
other cases, a prohormone
needing further processing)
41
Q

Additional modifications that may occur during translation in the ER lumen..

A

• Additional modifications that may occur during translation in the ER lumen
(contributing to the stability and folding of the final peptide hormone product):
• Covalent addition of oligosaccharide side chains (glycosylation)
• Folding and assembly of proteins, and quality control
(mediated by molecular chaperones in the ER)
• Disulfide bond formation

42
Q

Peptide/protein hormones – post-translational protein processing

Once ___ is terminated and proper folding and assembly have occurred, the
___ or ___ transferred to ____ (membranous regions)

A

Once translation is terminated and proper folding and assembly have occurred, the
hormone or prohormone is transferred to Golgi apparatus (membranous regions)

43
Q

Peptide/protein hormones – post-translational protein processing

• In the ____ further processing may incl…..

A 
• In the Golgi apparatus, further
processing may incl.:
proteolytic cleavage,
glycosylation/modifications of
carbohydrates, acylation,
acetylation, phosphorylation
44
Q

Peptide/protein hormones – post-translational protein processing

• Incorporated into ____ (membrane vesicles),
often as inactive ____:
cleavage by a ____ needed to
produce active hormone

A 
• Incorporated into secretory
granules (membrane vesicles),
often as inactive prohormone:
cleavage by a prohormone
convertase (PC) needed to
produce active hormone
45
Q

insulin post-translational processing

Preproinsulin

A

Preproinsulin: 110-a.a.,
biologically inactive.
In ER.

46
Q

insulin post-translational processing

Proinsulin:

A 
Proinsulin: 86-a.a.,
biologically inactive.
• In ER: folding,
disulphide bonds
• In Golgi apparatus: further
modifications (glycosylation)
47
Q

insulin post-translational processing

Insulin

A

Insulin: 51-a.a.,
biologically active as monomer.
In secretory granules.

48
Q

glucagon/GLP-1 & GLP-2 post-translational processing

____ is processed in a
tissue-specific manner,
depending in part on the local
dominant ____

A 
Proglucagon is processed in a
tissue-specific manner,
depending in part on the local
dominant prohormone
convertase (PC)
49
Q

glucagon/GLP-1 & GLP-2 post-translational processing

• In pancreatic α cells ,___
is synthesizes

A 
• In pancreatic α cells, glucagon
is synthesized (“opposes”
insulin: raises blood glucose)
50
Q

glucagon/GLP-1 & GLP-2 post-translational processing

• ___ cells synthesize
glucagon-like peptides (GLP)
• GLP-1 is what?

A 
• Enteroendocrine cells synthesize
glucagon-like peptides (GLP)
• GLP-1: a peptide hormone
which amplifies glucosestimulated insulin secretion
(among other effects)
51
Q

Post-translational modifications in the ER and Golgi apparatus
regulates….

A

Post-translational modifications in the ER and Golgi apparatus
regulates the synthesis of (typically tissue-specific) hormone variants

52
Q

Cytoskeletal protein-mediated migration of vesicles

A

Peptide hormones: released from secretory granules by exocytosis
• Cytoskeletal protein-mediated migration of vesicles toward cell surface,
vesicle (granule) fusion with the plasma membrane,
expulsion of contents (e.g., hormones) into the extracellular space

53
Q

Sorting

A

: Specific proteins meant to be secreted are
concentrated into secretory granules at the far side of
the Golgi apparatus. Sorting receptors that ”gather”
specific vesicle cargo may help mediate this.

54
Q

Budding

A

: Proteins that coat membrane-bound
transport vesicles gather and cause vesicle to break
free. These coat proteins mediate transfer of vesicles
within the cell.

55
Q

Trafficking

A

vesicles moved via motor proteins along

cytoskeleton (e.g. microtubules); requires energy.

56
Q

Docking/fusion

A

mediated by interactions with docking

proteins at the destination site

57
Q

Exocytosis is regulated.

A

Often triggered by changes in calcium concentrations in the cytoplasm
that affects secretory granule fusion
• Not continuous secretion – instead, acutely, in response to a stimulus
• Allows for rapid release of lots of hormone, vs. if hormone needed to be
synthesized de novo in response to the secretory signal
• The stimulatory factors may include metabolites, other hormones or
neuropeptides (which are released in response to detection of a changed variable
that affects homeostasis, for instance), or direct nerve innervation

58
Q

feedback loops

Input

A

Input: change in
extracellular
environment
or innervation

Only peripheral
endocrine glands
involved
(no CNS)

59
Q

Tropic hormone

A

a hormone
that cause the release of
another hormone

60
Q

pulsatile release

A

Hormone secretion = pulsatile release, may be important for physiological function
(rapid increase and subsequent decrease in hormone levels)
• Pulses may have variable frequency (from 4-30 min to 45-180 min) and
amplitude (e.g., up to 1000-fold changes for pituitary hormones)

  • insulin release
61
Q

secretion is pulsatile, and may be rhythmic

rhythmic changes

A

• Hormones may also show rhythmic changes – e.g., bursts every hour, every 24 h
(“circadian”) or less, or even longer
• Could be determined by environmental stimuli (e.g., light-dark cycle)
or an internal biological clock

62
Q

secretion is pulsatile, and may be rhythmic

Adrenocorticotropic hormone (ACTH)

A

: Adrenocorticotropic hormone (ACTH) has a notable circadian rhythm,
characterized by high levels during early morning hours

63
Q

ACTH

A

Adrenocorticotropic hormone (ACTH)

64
Q

secretion is pulsatile, and may be rhythmic

gonadotropins

A

follicle-stimulating hormone (FSH) and luteinizing

hormone (LH) have peaks every ~30 days (during menstrual cycle)

65
Q

Most peptide hormones: are …

A

soluble in aqueous
solutions (hydrophilic); therefore do not require
blood binding proteins for transport in the blood
stream

This makes them vulnerable to rapid degradation,
resulting in a relatively short plasma half-life and
duration of action

66
Q

Hormones in plasma: either ___ or ___ e.g. blood binding proteins

A

Hormones in plasma: either free form
(= biologically active form) or bound to other
molecules, e.g. blood binding proteins

67
Q

Blood binding proteins affect the controlled release

and stability of the pool of hormones

A

• Provides reservoir of hormone that exchanges
with free hormone fraction (making hormone
pool more stable, less dependent on
synthesis/release)

  • More uniform/distant distribution
  • Slowing hormone metabolism/breakdown

Not limited to hydrophobic hormones.
e.g., Growth Hormone and Insulin-like growth factor
1 are proteins with specific blood binding proteins.

68
Q 
Growth Hormone (GH)-Insulin-like
growth factor 1 (IGF-1) Axis:
A

• Regulates linear growth (growth in
height) in children/adolescents

  • GH
  • IGF-1
69
Q

• Growth Hormone (GH)-Insulin-like
growth factor 1 (IGF-1) Axis:
GH

A 
191-a.a. polypeptide hormone
synthesized by anterior pituitary;
~ half in circulation is bound to
specific GH-binding proteins
(reduce oscillations, prolong half-life)
70
Q

Feedback regulation:

A

The bulk of acute adjustments
to hormone levels = changes to
hormone synthesis & secretion

71
Q

key points of control regulation of synthesis/release (6)

A

Presence of regulators & interactions between regulators and
DNA regulatory elements influence the rate of gene transcription

Other control points: transport of RNA from the nucleus to the
cytoplasm, and the regulation of mRNA stability

Alternative splicing regulates the synthesis of
(typically tissue-specific) hormone or receptor variants

Control point: formation
of initiation complex
during translation

Post-translational modifications in the ER and Golgi apparatus
regulates the synthesis of (typically tissue-specific) hormone variants

Control of secretory granule release is an
important point of control for peptide hormones

72
Q

Protein/peptide hormones are ___ after
being internalized by target cells (example) , or ___ by nontarget cells, or cleared/filtered by kidney. They
may also be ___by extracellular ___ . (example)

A

Protein/peptide hormones are degraded after
being internalized by target cells (e.g., receptormediated clearance, with hormone degraded in
intracellular lysosomes), or degraded by nontarget cells, or cleared/filtered by kidney. They
may also be degraded by extracellular proteases
(e.g., in the plasma).

73
Q

Many peptide hormones: circulate in ____
un-bound to blood binding proteins (some
exceptions: ____). Therefore, relatively rapid
degradation/short duration of action

A

• Many peptide hormones: circulate in free form,
un-bound to blood binding proteins (some
exceptions: GH, IGF-1). Therefore, relatively rapid
degradation/short duration of action

74
Q

___ of hormone clearance from circulation
varies, but half life of un-bound peptide
hormones is generally in the ___
• vs. ___ for catecholamines (e.g.,
norepinephrine, epinephrine), and ___ for steroid & thyroid hormones

A

Rates of hormone clearance from circulation
varies, but half life of un-bound peptide
hormones is generally in the order of minutes
• vs. seconds for catecholamines (e.g.,
norepinephrine, epinephrine), and
hours for steroid & thyroid hormones

75
Q

____ (e.g., levels of
specific binding proteins or proteases) also
affects ____ levels

A

Control of hormone degradation (e.g., levels of
specific binding proteins or proteases) also
affects circulating hormone levels

76
Q

GLP-1 and GLP-1 based treatments for type 2 diabeties

A

GLP-1: a peptide hormone
which amplifies glucosestimulated insulin secretion,
among other effects)

Example: drugs that inhibit the primary protease
that degrades GLP-1, leading to higher GLP-1 levels

77
Q

Key points of control of hormone levels in circulation:

important for healthy physiological function ie…..

A

Key points of control of hormone levels in circulation:
important for healthy physiological function (i.e., they
allow hormone levels to respond to environmental &
internal changes, and thus carry out their function of
communicating & coordinating responses in other
cells in the body)

78
Q

Key points of control of hormone levels in circulation:
important for healthy physiological function (i.e., they
allow hormone levels to respond to environmental &
internal changes, and thus carry out their function of
communicating & coordinating responses in other
cells in the body) BUT…..

A

these may also be control points that underlie
dysregulation of hormone levels, leading to
pathological effects.

79
Q

Main types of cell surface receptors

for peptide hormones

A

• G-protein-coupled receptors
(or “7-transmembrane receptors”)

• Receptor-enzymes, including:
• Growth factor receptors
(have tyrosine kinase domain)
• Guanylyl cyclase receptors
(have guanylyl cyclase domain)

Type I cytokine receptors
(associated with an accessory protein
with a tyrosine kinase domain)

80
Q

• Interactions between hormones and their receptors depends on:

A
  • Number of receptors
  • Affinity of the hormone for the receptor
  • Concentra^on of circula^ng hormone
81
Q

Regulation at the level of the receptors is also an important point of
endocrine function control,…

A

Increasing or decreasing receptor synthesis
• Internalization vs. cell membrane localization of cell surface receptors
• Desensitization of receptors (“uncoupled” from signal transduction pathway, due
to such mechanisms as phosphorylation of the receptor)

82
Q 
Cells with more receptors will have a
\_\_\_\_ , at a given
concentration of hormone.
By the same principle, higher concentration
of hormone will lead to \_\_\_.
In both cases, this is only true up to a
\_\_\_\_, where all available
receptors are bound to the ligand)
A 
Cells with more receptors will have a
greater response (more [HR]), at a given
concentration of hormone.
By the same principle, higher concentration
of hormone will lead to more [HR].

[HR] = concentration of hormone-receptor complex
(hormone bound to receptor)

In both cases, this is only true up to a
saturation point, where all available
receptors are bound to the ligand)

83
Q

KD

A

KD is the equilibrium dissociation constant that
conventionally defines the affinity of the hormone-receptor binding.

KD is the concentration of free hormone [H]
at which half of available receptors are occupied.

The lower the KD, the higher the affinity of the receptor for the hormone.

84
Q 
Cells with \_\_\_ receptors will
have a greater response, at a given
concentration of hormone (up to a
\_\_\_\_\_, where all available
receptors are bound to the ligand)
A 
Cells with higher-affinity receptors will
have a greater response, at a given
concentration of hormone (up to a
saturation point, where all available
receptors are bound to the ligand)
85
Q

Iodocyanopindolol (ICYP)

A

Iodocyanopindolol (ICYP) is a β-adrenoceptor antagonist. Its [125I]-radiolabeled
derivative has been used to map the distribution of β-adrenoceptors in the body

Biol 432 (8 decks)

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