Insulin Degrading Enzyme by hcj

VIEWS: 77 PAGES: 11

									   For
BCMB8010
History
• First found Degrading the B chain of Insulin
• Many names: insulinase, insulin-degrading
  enzyme, insulin protease, insulin proteinase,
  insulin-degrading neutral proteinase,
  insulin-specific protease, insulin-glucagon
  protease, metalloinsulinase; IDE; amyloid-
  degrading enzyme
        Diebetes
Basic Properties
                           Alzheimer’s disease
• 110000 mol Zn2+ requiring
  metalloproteinase with
  neutral optimum
• Degrades insulin,with a Km
  of 20nM
• Cleaves substrates with no
  conserved cleavage
  sequence
Biochemical Properties continued

   Activator:                                Inhibitor:
   Substrates                                Chelators
   ATP




http://www.angelzfunnyz.com/Pets/tabid/54/AlbumID/378-2/Default.aspx
Isolation and Purification

            rat skeletal muscle
            human red blood cells
            Drosophila Kc cells
Raw
materials   rat liver
            human placenta               s          A


                                    Ni   Ni   Ni   Ni   Ni   Ni
 Recombinant            His
 protein

                    IDE
Ways of Assay

  125I-insulin
                  Abz-GGFLRKHGQ-EDDnp




  IDE digestion         IDE digestion

  TCA incubation


  Increased radioactive        HPLC
  signal in TCA supernatant
Structural Properties
The catalytic site


      H         E
          X X       H
The substrate conformations



                      Bound with
                      enzyme




                       Natural
                       structure
The enzyme conformations
      R state   Rate Limiting Step!!   T-state
5. Proteolytic Fragments of Insulsin(IDE) Retain Substrate Binding but lose Allosteric
Regulation, Eun Suk Song at al, Biochemistry, 2006, 45: 15085-15091
6. from ATP inhibits insulin-degrading enzyme activity, M.C.Camberos at al. Exp Biol
Med Vol, 2001,226:334-341
7. Affinity Purification of Insulin-degrading Enzyme and its endogenous inhibitor from
Rat Liver. Wataru Ogawa at al., the Journal of Biological Chemistry, 1992, 267:1310-



          Thank you!!!
1316
8. Purification of Insulin-specifc Protease by Affinity Chromatography, W.C. Duckworth
at. al, PNAS, 1972, 69: 3698-3702.
9. degradation products of insulin generated by hepatocytes and by insulin Protease,
W.C. Duckworth at al, JBC, 1988, 263:1826-1833.
10. structure And no need to applause a new substrate recognition
               of human insulin-degrading enzyme reveal
mechanism, Y.Shen at al, nature, 2006, 433:870-874.
11. Enzyme target to latch on to, nature, Malcolm A. Leissring and Dennis J. shlkoe,
2006, vol 443:761-762
12. Insuin-degrading enzyme as a downstream target of insulin receptor signaling
cascade: implications for Alzheimer’s Disease Intervention, Lixia Zhao, at al, Journal
of Neuroscience, 2004, 24:11120-11126.
13. Insulin clearance in Obesity, M.E.V. Mora, at al, Journal of the American College
of Nutrition, 2003, 22:487-493.
14. Links between Alzheimer's Disease and Diabetes, Miao-Kun Sun and Daniel L.
Alkon ,Timely Top Med Cardiovasc Dis. 2006 Oct 1;10:E24
15. insulin, insulin-degrading enzyme and amyloid-β peptide in Alzheimer’s disease:
review and hypothesis, W.Q. Qiu at al, Neurobiology of Aging, 2006, 27: 190-198

								
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