We evaluated the ability of three enzymes--N-acetyl-beta-D-glucosaminidase (NAG; EC 3.2.1.30), alanine aminopeptidase (AAP; microsomal aminopeptidase, EC 3.4.11.2), and gamma-glutamyltransferase (GGT; EC 2.3.2.2)--and adenosine deaminase binding protein (ABP) in urine to predict or confirm renal-transplant rejection in patients treated with cyclosporine. We measured the enzymes daily during the...

A phage repertoire was constructed using antibody genes from the bone marrow and the spleen of a rabbit immunized with Cry1Ab toxin. Biopanning against either the Cry1Ab toxin or a domain II loop 3 synthetic peptide resulted in the identification of monoclonal antibodies in scFv format. They inhibited binding and toxicity of Cry1Ab toxin against Manduca sexta. Toxin overlay assays, using the sc...

CLINICALCHEMISTRY, Vol. 32, No. 5, 1986 913 with either substrate, but the observed activity was 55.7% lower with iralanine-4-nitroanilide, in contrast with what happens in sera of males and nonpregnant women. Evidently, i..-alanine-4-mtroanilide behaves like LCHA in this regard, probably reflecting the relative resistance of these substrates to the action of cystyl-aminopeptidase and perhaps o...

One of the challenges in the development of methionine aminopeptidase (MetAP) inhibitors as antibacterial and anticancer agents is to define the metal ion actually used by MetAP in vivo and to discover MetAP inhibitors that can inhibit the metalloform that is relevant in vivo. Two distinct classes of novel nonpeptidic MetAP inhibitors that are not only potent but also highly selective for eithe...

F4(+) enterotoxigenic Escherichia coli (ETEC) strains cause diarrheal disease in neonatal and post-weaned piglets. Several different host receptors for F4 fimbriae have been described, with porcine aminopeptidase N (APN) reported most recently. The FaeG subunit is essential for the binding of the three F4 variants to host cells. Here we show in both yeast two-hybrid and pulldown assays that APN...

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The amino acid sequence of a decapeptide with growth hormone-releasing activity, isolated from porcine hypothalami has been determined. The Edman dansyl procedure was used on the intact peptide and on fragments isolated from tryptic and papain digests. The sequence found was confirmed by the results of digestion with aminopeptidase M, leucine aminopeptidase, and carboxypeptidase A and B. On the...

Methionine aminopeptidase (MetAP) is a metalloprotease that removes the N-terminal methionine during protein synthesis. To assess the importance of the two MetAPs in Mycobacterium tuberculosis, we overexpressed and purified each of the MetAPs to near homogeneity and showed that both were active as MetAP enzymes in vitro. We screened a library of 175,000 compounds against MtMetAP1c and identifie...

PepCs isolated from lactic acid bacteria and bleomycin hydrolases of eukaryotic organisms are strict aminopeptidases which belong to the papain family of thiol peptidases. The structural basis of the enzymic specificity of the lactococcal PepC has been investigated by site-directed mutagenesis. The deletion of the C-terminal residue (Ala-435) abolished the aminopeptidase activity, whereas this ...

The yeast vacuolar protein aminopeptidase I (API) is synthesized as a cytosolic precursor that is transported to the vacuole by a nonclassical targeting mechanism. Recent genetic studies indicate that the biosynthetic pathway that transports API uses many of the same molecular components as the degradative autophagy pathway. This overlap coupled with both in vitro and in vivo analysis of API im...