The Kell protein of the common K2 phenotype is a catalytically active metalloprotease, whereas the rare Kell K1 antigen is inactive. Identification of novel substrates for the Kell protein.
The Kell blood group is a highly polymorphic system containing over 20 different antigens borne by the protein Kell, a 93-kDa type II glycoprotein that displays high sequence homology with members of the M13 family of zinc-dependent metalloproteases whose prototypical member is neprilysin. Kell K1 is an antigen expressed in 9% of the Caucasian population, characterized by a point mutation (T193M) of the Kell K2 antigen, and located within a putative N-glycosylation consensus sequence. Recently, a recombinant, non-physiological, soluble form of Kell was shown to cleave Big ET-3 to produce the mature vasoconstrictive peptide. To better characterize the enzymatic activity of the Kell protein and the possible differences introduced by antigenic point mutations affecting post-translational processing, the membrane-bound forms of the Kell K1 and Kell K2 antigens were expressed either in K562 cells, an erythroid cell line, or in HEK293 cells, a non-erythroid system, and their pharmacological profiles and enzymatic specificities toward synthetic and natural peptides were evaluated. Results presented herein reveal that the two antigens possess considerable differences in their enzymatic activities, although not in their trafficking pattern. Indeed, although both antigens are expressed at the cell surface, Kell K1 protein is shown to be inactive, whereas the Kell K2 antigen binds neprilysin inhibitory compounds such as phosphoramidon and thiorphan with high affinity, cleaves the precursors of the endothelin peptides, and inactivates members of the tachykinin family with enzymatic properties resembling those of other members of the M13 family of metalloproteases to which it belongs.
Clapéron
Audrey
A
INSERM U573, 75014 Paris, France.
Rose
Christiane
C
Gane
Pierre
P
Collec
Emmanuel
E
Bertrand
Olivier
O
Ouimet
Tanja
T
D000941:Antigens / Q000737:chemistry; D000954:Antigens, Surface / Q000737:chemistry / Q000502:physiology*; D001798:Blood Proteins / Q000737:chemistry / Q000502:physiology*; D001854:Bone Marrow Cells / Q000378:metabolism; D020126:Brefeldin A / Q000494:pharmacology; D002384:Catalysis; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002469:Cell Separation; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000378:metabolism; D005434:Flow Cytometry; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D006868:Hydrolysis; D020014:K562 Cells; D007700:Kinetics; D013058:Mass Spectrometry; D008666:Metalloendopeptidases / Q000737:chemistry; D045726:Metalloproteases / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D015288:Neurokinin A / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D010641:Phenotype; D016133:Polymerase Chain Reaction; D021241:Spectrometry, Mass, Electrospray Ionization; D013379:Substrate Specificity; D015320:Tachykinins / Q000737:chemistry / Q000378:metabolism; D013696:Temperature; D014162:Transfection; D015032:Zinc / Q000737:chemistry
D000941:Antigens; D000954:Antigens, Surface; D001798:Blood Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D010455:Peptides; D015320:Tachykinins; C041106:kell-active proteins; D020126:Brefeldin A; D015288:Neurokinin A; D006026:Glycoside Hydrolases; D045726:Metalloproteases; D008666:Metalloendopeptidases; D015032:Zinc