Secretion of human furin into mouse milk.
We have previously described the expression of the human proprotein convertase furin or paired basic amino acid-cleaving enzyme, in mice transgenic for paired basic amino acid-cleaving enzyme and human Protein C (HPC). Here we show 100-fold or higher expression of furin in the mammary gland, compared with endogenous furin. Furin and recombinant HPC were detected in the same regions of the mammary gland and regulated similar to the endogenous whey acidic protein. In addition to the expected intracellular localization, furin was secreted into the milk as an 80-kDa form lacking the transmembrane and cytoplasmic domains. Furin present at levels of up to 40,000 units/ml milk cleaved the t-butoxycarbonyl-RVRR-AMC substrate with a Km of 32 microM, and processed the recombinant HPC precursor at the appropriate sites. Surprisingly, the expression of an active protease was not toxic to the mammary gland. This is a rare example of an animal model secreting active truncated forms of a processing endoprotease into a bodily fluid.
Paleyanda
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RK
J. Holland Laboratory, American Red Cross, Rockville, Maryland 20855, USA.
Drews
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R
Lee
T K
TK
Luboń
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H
D000818:Animals; D045683:Furin; D006801:Humans; D007150:Immunohistochemistry; D008321:Mammary Glands, Animal / Q000737:chemistry*; D051379:Mice; D008892:Milk / Q000737:chemistry*; D008894:Milk Proteins / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013381:Subtilisins / Q000235:genetics / Q000378:metabolism*; D019076:Transgenes
D008894:Milk Proteins; D011993:Recombinant Fusion Proteins; C024471:whey acidic proteins; D013381:Subtilisins; D045683:Furin