No Slide Title · comprendono sfingomieline, cerebrosidi e gangliosidi. c) Oligosaccaridosi:...
Transcript of No Slide Title · comprendono sfingomieline, cerebrosidi e gangliosidi. c) Oligosaccaridosi:...
Proteoglycans Are Assembled in the Golgi Apparatus
Modificazioni post-traduzionali delle proteine
La O-glicosilazione
La O-glicosilazione è un processo altamente specifico, che non vede l'aggiunta "seriale" di
carboidrati alla proteina in processazione.
Si svolge completamente nell'apparato del Golgi, dove zuccheri vengono legati al
peptide a livello dell'atomo di ossigeno delle catene laterali di serina o treonina.
L'aggiunta riguarda un singolo carboidrato alla volta e solitamente il numero di zuccheri
legati durante questo processo è limitato a pochi residui.16
Serina o
treonina
Affected gene Protein Affected PTM Disease Major clinical manifestations
MAN1B1 α 1,2 mannosidase glycosylation MAN1B1-CDG severe mental retardation, delayed speech
SLC35A1 CMP-sialic acid transporter glycosylation SLC35A1-CDG seizures, intellectual disability, ataxia, bleeding
SLC35A2 UDP-galactose transporter glycosylation SLC35A2-CDG intellectual disability, seizures, skeletal abnormalities
SLC35A3 UDP-GlcNAc transporter glycosylation Arthrogryposis, mental retardation and seizures autism spectrum disorder, hypotonia, epilepsy, and arthrogryposis
SEC23B Sec23 homolog B glycosylation Dyserythropoietic anemia, congenital, type IIerythroblastic anemia: splenomegaly, gallstones, and iron overload potentially with liver cirrhosis or
cardiac failure.
TRIP11 Golgi microtubule associated protein 210 glycosylation Achondrogenesis type 1A severe chondrodysplasia, lethal before or shortly after birth
UBE3A Ubiquitin ligase E3A glycosylation Angelman syndrome intellectual disability, seizures, lack of speech, and characteristic abnormal behavior
COG2 Component of oligomeric Golgi complex 2 glycosylation COG2-CDGmicrocephaly, developmental delay, intellectual disability, seizures, facial dysmorphism, liver
dysfunction
SLC33A1Solute carrier family 33 (acetyl-CoA
transporter), member 1acetylation Spastic paraplegia-42 spastic gait, increased lower limb tone, weakness and atrophy of the lower limb muscles, pes cavus
CHST3 Chondroitin 6-O-sulfotransferase sulfationSpondylo-epiphyseal dysplasia with joint
dislocationsunusual skeletal dysplasia
CHST6Corneal N-acetylglucosamine-6-O-
sulfotransferasesulfation Macular corneal distrophy type II progressive corneal opacification and reduced corneal sensitivity
CHST8 GalNAc-4-O sulfotransferase I sulfation Peeling skin syndrome general skin peeling
CHST14Dermatan sulfate GalNAc-4-O
sulfotransferase Isulfation
Ehlers-Danlos syndrome musculocontractural
type 1craniofacial dysmorphism, congenital contractures of thumbs and fingers, clubfeet, severe kyphoscoliosis
ARSE Arylsulfatase E sulfation Chondrodysplasia punctata 1 stippled epiphyses, brachytelephalangy, nasomaxillary hypoplasia
PAPPS2 PAPS synthase sulfation Brachyolmia type 4short-trunk stature, rectangular vertebral bodies, precocious calcification of rib cartilages, short femoral
neck. Early death for severe cases.
SLA26A2 Sulfate anion transporter sulfation Achondrogenesis type 1B severe chondrodysplasia, early death of respiratory failure
Atelosteogenesis type 2 pulmonary hypoplasia, lethal in infants
Epiphyseal dysplasia multiple 4 joint pain, scoliosis, malformations of the hands, feet, and knees
Diastrophic dysplasia scoliosis, clubfeet, malformed pinnae with calcification of the cartilage, cleft palate in some cases
GNTPGN-acetylglucosamine-1-
phosphotransferase gamma subunitphosphorylation Mucolipidosis III gamma short stature, skeletal abnormalities, cardiomegaly, and developmental delay
GNTPABN-acetylglucosamine-1-
phosphotransferase alpha and beta subunitsphosphorylation Mucolipidosis II and III
Hip dislocation, gingival hyperplasian, thoracic deformities and hernia soon after birth. Delayed
psychomotor development. Same clinical features for mucolipidosis III as described just above.
IMPAD1 Golgi-resident PAP phosphatase phosphorylation Chondrodysplasia with joint dislocationsshort stature, chondrodysplasia with brachydactyly, congenital joint dislocations, micrognathia, cleft
palate, and facial dysmorphism
INPP5E Inositol polyphosphate-5-phosphatase phosphorylation Morm syndrome Mental retardation, truncal obesity, retinal dystrophy, and micropenis
phosphorylation Joubert syndrome 1Heterogenous: hypoplasia of the cerebellar vermis with the characteristic neuroradiologic molar tooth
sign , dysregulation of breathing pattern and developmental delay.
AKAP9 A-kinase anchor protein 9 phosphorylation Long QT syndrome-11 recurrent syncope, seizure, or sudden death
FAM20CGolgi kinase (family with sequence
similarity 20, member C)phosphorylation Raine syndrome neonatal osteosclerotic bone dysplasia, increased ossification of the skull
CAMKMT Calmodulin-lysine N-methyltransferase methylation 2p21 deletion syndrome cystinuria, neonatal seizures, hypotonia, severe somatic and developmental delay, facial dysmorphism
MBTPS2 Site-2 protease proteolytic cleavageIFAP syndrome with or without BRESHECK
syndromeichthyosis follicularis, atrichia, and photophobia
ZDHHC8 Zinc finger, DHHC-type containing 8 palmitoylation Schizophrenia susceptibilityhallucinations and delusions, inappropriate emotional responses, disordered thinking and concentration,
erratic behavior
ZDHHC9 Zinc finger, DHHC-type containing 9 palmitoylation X-linked mental retardation (Raymond type) general intellectual limitations associated with impairments in adaptive behavior
ZDHHC15 Zinc finger, DHHC-type containing 15 palmitoylation X-linked mental retardation-91 general intellectual limitations associated with impairments in adaptive behavior
PPT1 Palmitoyl-protein thioesterase 1 palmitoylation Neuronal ceroid lipofuscinosis 1Heterogenous: progressive dementia, seizures, and progressive visual deficiency. The cellular phenotype
includes intracellular accumulation of autofluorescent lipopigment storage material.
Degradazione delle macro molecole
EXTRACELLULARI
Funzioni dei lisosomi
(Proteasoma degrada le proteine citosoliche
Funzioni dei lisosomi
(Proteasoma degrada le proteine citosoliche INTRACELLULARI
Degradazione di organelli
Anatomia di un Lisosoma
LisosomiRecupero dei
prodotti
degradati
Mannose 6-phosphate (M6P)
sent to lysosomes
Mannose
How do lysosomal enzymes get to the lysosome?
lysosome
Biosynthesis and traffickingof a lysosomal hydrolase
receptor-dependenttransport
receptor recycling
binding to M6Preceptor
removal of phosphate
addition of phosphate
dissociation at acidic pH
addition of clathrin coat
mature lysosomal hydrolase
Golgi apparatus
Man-6P as a lysosomal targeting signal for proteins with N-linked glycosylation
D-Mannose
Mannose
Cytoplasm ER
Golgi
ASN Transferase absent in I-cell Disease
Receptor is on lysosome bound vesicles but will also end up on the cell surface
Classificate in vari gruppi:
a) Mucopolisaccaridosi: difetto nella degradazione dei mucopolisaccaridi (o GAG),
che svolgono importanti funzioni nel tessuto connettivo.
b) Sfingolipidosi: difetto nella degradazione lisosomiale degli sfingolipidi, che
comprendono sfingomieline, cerebrosidi e
gangliosidi.
c) Oligosaccaridosi: accumulo di oligosaccaridi e di glicoproteine. In
questa classe rientrano la Fucosidosi, la Sialidosi, la Mucolipidosi,
la Mannosidosi.
- Malattie dovute a trasporto lisosomiale alterato.
- Malattie dovute al mancato trasporto degli enzimi lisosomiali.
- Altre malattie lisosomiali (es. Malattia di Pompe, dovuta ad un
deficit di a-glucosidasi).
Malattia
Materiale prevalentemente
accumulato Deficit enzimatico
M. di Gaucher Glucorebroside Glucorebrosidasi
M. di Niemann-Pick Sfingomielina Sfingomielinasi
Leucodistrofia globoide Galattocerebroside Galattocerebrosidasi
Leucodistrofia metacromatica Sulfatide Aril-sulfatasi A
M. Di Fabry Globotriaosilceramide Alfa-galttosidasi A
Fucosidosi Pentaesosilfuco-glicolipide Alfa-fucosidasi
M. Di Farber Ceramide Ceramidasi
Gangliosidosi generalizzata Ganglioside GM1GM1 ganglioside: beta
galattosidasi
M. Di Tay-Sachs Ganglioside GM2 Esosaminidasi A
M. Di SandhoffGanglioside GM2
Esosaminidasi A e B
I-Cell disease (Mucolipidosis Type II)
Autosomal recessive (1/640,000 live birth)
Absence of N-acetylglucosaminyl-1-phosphotransferase.
No Man-6P signal -> enzymes are not targeted to lysosomes -> they are instead secreted to the media
Build up of proteins which cannot be degraded in vacuols. Massive increase in number and size of vacuols which
compromises cell architecture and function.
Overall symptoms
Growth failure and failure to thrive are rapidly progressive.
Developmental delay is severe and often the presenting symptom.
Coarse facial features and musculoskeletal abnormalities
Frequent upper respiratory tract infections
Death from pneumonia or congestive heart failure occurs in early childhood.
I-cell disease patient, 3.5 years old, with her 1week-old sister
Tiny size (not much bigger than her newborn sister)Skeletal abnormalities causing disproportionately large headFingers clenched (joints can’t relax)Gums very swollen
The patient died shortly after this picture was taken.
lysosome
Biosynthesis and traffickingof a lysosomal hydrolase
receptor-dependenttransport
receptor recycling
binding to M6Preceptor
removal of phosphate
addition of phosphate
dissociation at acidic pH
addition of clathrin coat
mature lysosomal hydrolase
Golgi apparatus
Fabry
• Fabry disease is a lysosomal storage disorder seen in one out of every 40,000
• It is caused by a deficiency in the enzyme alpha-galactosidase
• which then results in the body’s inability to break down specific fatty substances called globotriaosylceramide (abbreviated GL-3 or Gb3).
Hurler- Schie
• Mucopolysaccharidosis I is a lysosomal storage disorder that is abbreviated MPS I and sometimes called Hurler syndrome, Hurler-Scheie syndrome, or Scheie syndrome.
• It is caused by a deficiency in the enzyme alpha-iduronidase
• which is needed to break down certain complex sugars called glycosaminoglycans (abbreviated GAGs and formerly called mucopolysaccharides).
MPS IV (sindrome di Morquio)
MPS VI (sindrome di Maroteaux-Lamy)
ENDOCITOSITrasporto vescicolare
Materiale extracellulare è trasportato
all’interno della cellula in vescicole
di endocitosi, rivestite di clatrina,
che gemmano dalla membrana
plasmatica e si fondono con gli
endosomi precoci.
Al livello degli endosomi precoci, i
componenti di membrana che hanno
preso parte al processo , vengono
riciclati e fanno ritorno alla
membrana plasmatica, mentre gli
endosomi precoci si trasformano
gradualmente in endosomi tardivi.
L’endocitosi definisce l’insieme dei processi di
trasporto intracellulare di sostanze o organismi
extracellulari
Tre tipi di endocitosi:
1. Pinocitosi
2. Fagocitosi
3. Endocitosi mediata da recettore
In genere il denominatore comune di questi meccanismi di
internalizzazione di sostanze extracellulari è la formazione
di vescicole più o meno voluminose delimitate da
membrana
Le sostanze raggiungono i
lisosomi attraverso 4 vie
• Pinocitosi generalizzata
– Cellula “beve” < 150 nm
• Endocitosi Mediata da Recettore
• Fagocitosi
– Cellula “mangia” > 250 nm
• Autofagia
– Cellula “mangia se stessa” organelli invecchiati o non
più funzionali
– Meccanismo importante nello sviluppo, nell’omeostasi
e nella morte cellulare
Sostanze disciolte in acqua (soluti) sono continuamente
internalizzate dalle cellule via Pinocitosi.
Piccole invaginazioni della membrana plasmatica formano
microvescicole conteneti i soluti che progressivamente
vengono rilasciati nel citoplasma
Endocitosi
• Pinocitosi
– Letteralmente: la cellula beve
– Importo di fluidi che vengono
rilasciati nel citosol
Pinocytosis(a form of endocytosis)
2. FAGOCITOSI
FAGOCITOSI
Escape of Listeria monocytogenes from a Vacuole
Autophagy Degrades Unwanted Proteins and Organelles
Autophagy Degrades Unwanted Proteins and Organelles
Autophagy Degrades Unwanted Proteins and Organelles
clathrin coated
pitvesicle
IN
OUT
IN
OUT
IN= cytosol
3. receptor mediated endocytosis
adaptin
cargo receptor
OUT
IN
clathrin
OUT
IN
+
dynamin
OUT
IN
clathrin coated vesicle
OUT
IN
GTP GDP + Pi
pinching off
uncoating
ATP ADP + Pi
clathrin coated vesicle
OUT
IN
cytosol
v-SNAREt-SNARE
docking
OUT
IN
Endosome
fusion
Receptor-mediated endocytosisEndosome
cytosol
OUT
clathrin triskelions clathrin cage
Michael Brown
Joseph Goldstein
Noble Prize 1985
"for their discoveries concerning the
regulation of cholesterol metabolism"
LDL
= low density lipoprotein
cholesterol
out
cytosol
Receptor recyclingout
pH: 5.5
out
cytosol
Hypercholesterolemia
Atherosclerosis
The mutations
• Five classes
– I – can’t synthesize LDLR
– II – Targeting mutation
– III – can’t bind LDL
– IV – can’t cluster receptor in clathrin coated pit and internalize
– V – can’t recycle LDL
EarlyEndosome
Transportvesicle
Recyclingvesicle
Microtubule
Endosomecarrier vesicle Dynein
Degradation of the Ligand and Recycling of its Receptor
COP-I
LDL LDL receptor
Actin
Recyclingvesicle
EarlyEndosome
Transportvesicle
Microtubule
Endosomecarrier vesicle Dynein
Fe
Recycling of the Ligand and its Receptor
Transferrin-Fe Transferrin
Actin
Receptor
EarlyEndosome
Transportvesicle
Recyclingvesicle
Microtubule
Endosomecarrier vesicle Dynein
Transcytosis of the Ligand and its Receptor
IgG
Many Proteins and Lipids Are Carried Automatically from the Trans Golgi Network (TGN) to
the Cell Surface
Secretory Vesicles Bud from the Trans Golgi Network
secretion
retrovirus
15.9
Viral env