Neprilysin-Like Protease, alpha (NEPL-a)

The amyloid beta peptide (Ab) of 39-43aa is constitutively produced in brain upon proteolysis of the b-amyloid precursor protein (APP) and exists as fragments of 40, 42 and 43aa (Ab40, Ab42, Ab43). In young and healthy humans, Ab is rapidly catabolized before it can be deposited in the brain. Upon aging or the onset of familial Alzheimer's Disease however, alterations in either synthesis or degradation/ clearance of Ab may contribute to amyloid depositions in the brain. Ab is susceptible to a number of in vivo and in vitro proteases like cathepsin-D and M-13 metalloproteases. The M-13 family comprises several zinc-dependent metalloproteases like DINE, PHEX, KELL, ECE, XCE, neprilysin (NEP) and neprilysin-like proteases (NEPLs). The NEPLs (NEPL-alpha, NEPL-beta, NEPL-gamma) arise from the alternative splicing of a single NEPL gene and are zinc dependent metalloproteases with ~54 % homology to NEP. NEPL-a is a type II trans-membrane metallopeptidase (~100kD non-glycosylated and 120kD glycosylated) containing a highly glycosylated polypeptide chain of 742aa with a HEXXH zinc-binding consensus, cytoplasmic tail and a large extracellular catalytic domain. NPL-a lacks a 23aa sequence present in N-terminal of NEPL-b. In contrast to NEP and NEPL-b which exist in membrane-bound as well as soluble forms, NEPL-a is expressed as membrane-bound form only. In comparison to NEP, the NEPL-a has lower affinity and/or smaller Vmax for Ab and cannot proteolyze cell secreted Ab40 or Ab42 in vivo. Unlike NEP, NEPL-a is not a major in vivo peptidase for degrading endogenous Ab however, it might degrade Ab in Alzheimer patients where Ab is accumulated in excess and an Ab-degradative pathway, alternative to NEP, exists.\nApplications: \nSuitable for use in ELISA and Western Blot Other applications not tested.\n\nRecommended Dilutions:\nELISA: 1:5000-1:50,000 using 50-100ng of control peptide (N2030-34) per well.\nWestern Blot: 1:1000-1:5000 using ECL. \nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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SPECIFICATIONS

Catalog Number

N2030-30

Size

50ul

Applications

ELISA, WB

Hosts

Rabbit

Reactivities

Mouse

Form

Supplied as a liquid, neat serum, 40% glycerol.

P Type

Pab

Purity

Serum

Isotype

IgG

References

(1) Shirotani K et al (2001) JBC 276, 21895-21901; Iwata et al. (2001) Science 292, 1550; Ikeda et al. (1999) J. Biol. Chem. 274, 32469; Boileau et al. (2001) Biochem. J 355, 107; Kiryu-Seo et al. (2000) PNAS 97, 4345.\n

Additional Info

Recognizes mouse Neprilysin-Like Protease, alpha (NEPL-a).