tailieunhanh - Báo cáo khoa học: The role of ADAM10 and ADAM17 in the ectodomain shedding of angiotensin converting enzyme and the amyloid precursor protein

Numerous transmembrane proteins, including the blood pressure regulating angiotensin converting enzyme (ACE) and the Alzheimer’s disease amyloid precursor protein (APP), are proteolytically shed from the plasma membrane by metalloproteases. We have used an antisense oligo-nucleotide (ASO) approach to delineate the role of ADAM10 and tumour necrosis factor-aconverting enzyme (TACE;ADAM17) in theectodomainsheddingofACEand APP from human SH-SY5Y cells. | Eur. J. Biochem. 271 2539-2547 2004 FEBS 2004 doi The role of ADAM10 and ADAM17 in the ectodomain shedding of angiotensin converting enzyme and the amyloid precursor protein Tobias M. J. Allinson1 Edward T. Parkin1 Thomas P. Condon2 Sylva L. U. Schwager3 Edward D. Sturrock3 Anthony J. Turner1 and Nigel M. Hooper1 1 Proteolysis Research Group School of Biochemistry and Microbiology University of Leeds UK 2Isis Pharmaceuticals Carlsbad CA USA 3Division of Medical Biochemistry University of Cape Town South Africa Numerous transmembrane proteins including the blood pressure regulating angiotensin converting enzyme ACE and the Alzheimer s disease amyloid precursor protein APP are proteolytically shed from the plasma membrane by metalloproteases. We have used an antisense oligonucleotide ASO approach to delineate the role of ADAM10 and tumour necrosis factor-a converting enzyme TACE ADAM17 in the ectodomain shedding of ACE and APP from human SH-SY5Y cells. Although the ADAM10 ASO and TACE ASO significantly reduced 81 their respective mRNA levels and reduced the a-secretase shedding of APP by 60 and 30 respectively neither ASO reduced the shedding of ACE. The mercurial compound 4-aminophenylmercuric acetate APMA stimulated the shedding of ACE but not of APP. The APMA-stimulated secretase cleaved ACE at the same Arg-Ser bond in the juxtamembrane stalk as the constitutive secretase but was more sensitive to inhibition by a hydroxamate-based compound. The APMA-activated shedding of ACE was not reduced by the ADAM10 or TACE ASOs. These results indicate that neither ADAM10 nor TACE are involved in the shedding of ACE and that APMA which activates a distinct ACE secretase is the first pharmacological agent to distinguish between the shedding of ACE and APP. Keywords ADAM antisense oligonucleotide metalloprotease secretase tumour necrosis factor-a converting enzyme. Angiotensin converting enzyme ACE is critically involved in blood pressure .