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CSB327 > MMPs > Flashcards

MMPs Flashcards

1
Q

What can be used as a loading control for western blot and why?

A

Beta actin.

commonly chosen as a loading control due to its general expression across all eukaryotic cell types. The expression levels of this protein do not vary drastically due to cellular treatment, which is another reason the protein makes a suitable control.

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2
Q

What is the inhibitor of MMP? Which types inhibit which types of MMP?

A

TIMP
1 - MMP2, 9, 13
2,3,4 - MT-MMP

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3
Q

When is MT-MMP expression the highest?

A 
Embryogenesis
ECM remodelling (wound healing, angiogenesis, cancer)
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4
Q

What do inactive proMMPs look like? How are they activated?

A

pro domain’s SH in cysteine binds with zinc in catalytic domain.
Zn2+ binding endopeptidase cleaves the pro domain

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5
Q

Where is proMMP cleaved?

A

in the pericellular environment

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6
Q

What are the substrate specific chacteristics in the secreted MMPs?

A

catalytic FN type II repeats

hemopexin domain

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7
Q

Characteristic of the catalytic domain in secreted MMPs?

A

Zn2+

FN type II repeats

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8
Q

FN type II is required for cleaving what?

A

elastins, gelatins, collagen IV,

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9
Q

Hemopexin is essential for cleaving what?

A

collagens

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10
Q

Whats the ratio of TIMP and MMP bind?

A

1:1

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11
Q

Explain the process of wound healing

A
  1. keratinocyte migration over basal lamina
  2. provisional matrix (fibrin, plasma protein) is removed
  3. fibroblast contracts the ECM to close the wound
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12
Q

Some ECM and non ECM molecules turned by MT1-MMP

A

ECM

  • collagen I, II, III
  • fibronectin
  • laminin
  • perlecan
  • aggrecan
  • syndecan 1 and 4

non ECM

  • CD44
  • Pro TNF alpha
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13
Q

Fibronect turnover is done by which MMP?

A

MT1-MMP

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14
Q

which Integrin is regulated by MT1-MMP for fibronectin turnover?

A

a5b1

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15
Q

Which molecule’s shedding activity is linked to tumor progression? and which MMP is responsible for this?

A

syndecans 1 and 4

MT1-MMP

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16
Q

Where is MT-MMP pro domain cleaved?

A

trans golgi

17
Q

What are some differences in structure of soluble and MT MMP?

A

MT-MMP has

  • furin recognition motif
  • transmembrane domain
  • Glycosyl phosphotidylinositol (GPI) anchor
18
Q

Describe the activation cascade of MT1-MMP

A
  1. pro domain cleaved in trans golgi
  2. dimer presenting at the cell surface
  3. one MT1-MMP is inactivated by TIMP-2
  4. Other MT1-MMP cleaves pro domain in MMP2 activating it. It can also activate MMP13
  5. MMP2 and 13 activates MMP9
19
Q

which domain allows MT1-MMP dimerization?

A

pex domain

20
Q

What is the purpose of the furin domain?

A

allows trans golgi cleavage of MT1-MMP instead of extracellular

21
Q

How are only half of the MT1-MMPs inactivated?

A

1:2 ratio of TIMP2:MT1-MMP

22
Q

What do disintegrins bind do, via what?

A
  • binds to integrins via XCD (ADAM 15 –RGD)

- also fibronectin

23
Q

Venom from viper has what?

A

disintegrins

24
Q

which ADAMs family binds to integrin a5b1?

A

ADAM-15 and -17

25
Q

Differences between ADAMs and MT1-MMP

A
  • disintegrins
26
Q

How is ADAMs pro domain removed?

A

furin like convertase or by autocatalysis

27
Q

Differences between ADAMs and ADAM-TS

A
  • Thrombospodin motif

- ADAM-TS is secreted

28
Q

main function of ADAMs

A

shedding extracellular domains of transmembrane proteins

29
Q

ADAM

A

ADAMTS-4 (aggrecanase-2) – aggrecan turnover

30
Q

ADAMTS-4 mainly turns over what?

A

aggrecan

31
Q

which disease is associated with ADAMTS-4?

A

arthritis due to excessive aggrecan turnover

32
Q

Other than MT1-MMP what can activate pro-MMP2?

A

ROS, TBI

33
Q

what type of barriers are in place in BBB?

A

classical tight junctions

34
Q

Functions of BBB?

A
  • Allows diffusion of O2 via transporters
  • nutrients, Hormones, etc.
  • Provides protection against pathogens and toxic substances
35
Q

What happens when tight junctions are broken in BBB?

A

plasma protein leakage

36
Q

What are some immune responses to broken BBB junctions?

A
  • leukocytes
  • more fibrinogen
  • activate microgilla
  • MMP activity increase > ECM degredation
  • activated cytokines e.g. IL-1β and TNF-a
37
Q

How does viper venom work?

A

Disintegrin binds to intrgrins and prevents early step in blood clotting. Usually through the ADAM 15-RGD

38
Q

which MMP activity is associated with arthritis?

A

ADAM-TS 4

CSB327 (5 decks)

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