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Regulation of protein phosphorylation by insulin/IGF-1. Yang,Yu-Ying Tseng, Yu-Hua C.Ronald Kahn. Insulin. Insulin

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regulation of protein phosphorylation by insulin igf 1

Regulation of protein phosphorylation by insulin/IGF-1

Yang,Yu-Ying Tseng, Yu-Hua C.Ronald Kahn

insulin
Insulin
  • Insulin

an anabolic hormone with strong metabolic effects. If concentration of glucose is too high, then insulin will be released to metabolize the glucose. When it binds to its receptor, the receptor will autophosphorylation, activate a family of IRS proteins, and lead the transmitting the signal downstream.

igf 1
IGF-1
  • IGF-1

insulin-like growth factor 1,has it’s own receptor.It has a similar signal transduction pathways to insulin’s. IGF-I mediates many actions of growth hormone, and it stimulates cell replication, cell differentiation and the synthesis of cellular products. As for their biological effects, in general, IGF-1 is mainly responsible for mitogenic effects

irs insulin receptor substrate proteins
IRS(insulin receptor substrate)proteins
  • IRS-1

its deficiency causes growth retardation and insulin resistance, which will lead to type 2 diabetes.

  • IRS-2

insulin resistance, lead to serious diabetes when it absents

slide6
IRS-3

no glucose metabolism or growth problems when it is absent

  • IRS-4

slight defects in growth, reproduction, and glucose homoeostasis

method
Method
  • brown pre-adipocytes from different IRS KO mice
  • Western blotting

-- sample preparation

cell + <insulin/IGF-1> for <5mins/30mins>

scrap cell

normalize cell

-- loading gel

-- transfer

-- immunoblotting

-- ECL

creb p insulin 100 nm

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IRS-3 KO

m1

IRS-3 KO

m2

IRS-1,3 KO

Wt

IRS-1 KO

IRS-2 KO

Creb-pInsulin (100 nm)

ST5

(min)

CREB-p

ATF-p

IRS-1 KO

+ IRS-4

IRS-1 KO

+ IRS-1

IRS-1 KO + IRS-3

IRS-1 KO

+ IRS-2

Wt

IRS-1 KO

ST6

(min)

CREB-p

ATF-p

creb p igf 1 100nm
Creb-p (IGF-1 100nm)

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IRS-3 KO

m2

IRS-3 KO

m1

IRS-1,3 KO

Wt

IRS-1 KO

IRS-2 KO

ST5

(min)

CREB-p

ATF-p

IRS-1 KO

+ IRS-3

IRS-1 KO

+ IRS-4

IRS-1 KO

+ IRS-1

IRS-1 KO

+ IRS-2

Wt

IRS-1 KO

ST6

(min)

CREB-p

ATF-p

akt juk
Akt-Juk

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IGF-1 (100 nm)

P85a KO

+ P85a

IRS-1 KO

+ IRS-1

IRS-3 KO

Wt

IRS-1 KO

IRS-2 KO

IRS-1,3 KO

P85a KO

ST 4

(min)

Akt

Jnk

Insulin (100 nm)

P85a KO

+ P85a

IRS-1 KO

+ IRS-1

ST 4

IRS-3 KO

Wt

IRS-1 KO

IRS-2 KO

IRS-1,3 KO

P85a KO

(min)

Akt

Jnk

result
Result
  • Insulin and IGF-1 almost have the same stimulative efficiency.
  • Compare with Wt cell, the KO cells did have lower efficiency in signaling.
  • IRS1+3KO cells didn’t lower much efficiency in signaling.
  • Akt-Juk shows stronger bands than Creb-p in the same time period stimulating.
conclusion
Conclusion
  • both insulin and IGF-1 can cross-react with each other\'s receptor when used at high dosages.
  • Using differentiation protocols, both with and without insulin, preadipocyte cell lines derived from IRS-1 KO mice exhibited a marked decrease in differentiation and protein

accumulation (10 to 40%) compared to wild-type cells (90 to 100%).

slide13
IRS-1KO gene maybe complementary to IRS-2, IRS-3 gene, but not IRS-4 gene.
  • IRS-1KO and IRS-3 KO cell didn’t result in much deficiency in signaling, maybe there’s other pathways for the insulin/IGF-1 signaling.
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