Molecular cloning of soluble aminopeptidases from Saccharomyces cerevisiae

Abstract

Plasmids capable of complementing lupl, lap2 and lap3 mutations [R. J. Trumbly and G. Bradley (1983) J. Bucteriol. 156, [36][37][38][39][40][41][42][43][44][45][46][47][48] were isolated from a yeast YEpl3 library by screening for activity against the chromogenic aminopeptidase substrate L-leucine P-naphthylamide in intact yeast colonies. The genomic inserts were shown to contain the structural genes for aminopeptidases yscII, yscIII and ysclV. Plasinids containing the gene encoding aminopeptidase yscII of Sacchuromyce.y cerevisiae, APE2 (LAPI) were analyzed in detail. APE2 was determined by DNA blot analysis to be a singlecopy gene located on chromosome XI. The cloned fragment was used to identify a 2.7-kb mRNA. The proteolytic system of the yeast Succhuromyces cerrvihiue has attracted considerable attention and is still actively studied (for reviews, see [I, 21). The yeast proteinases located in the vacuole, the lysosome-like organelle [3], are by far the best characterized. The genes encoding proteinase yscA, proteinase yscB, aminopeptidase yscI, dipeptidyl aminopeptidase yscV, carboxypeptidase yscY and carboxypeptidase yscS have been cloned and sequenced [4-221 and their biosynthesis and sorting have been examined (reviewed in [13]).Matile et al. [14] detected four aminopeptidases with activity on leucine /3-naphthylamide after separating crude extracts from yeast by starch gel electrophoresis. Four aminopeptidases capable of hydrolyzing tripeptides were deCorrespondeelwe to P. SuCrez-Rendueles,

Saccharomyces boulardii Enhances N-Terminal Peptide Hydrolysis in Suckling Rat Small Intestine by Endoluminal Release of a Zinc-Binding Metalloprotease

“…In addition, the enzymatic reaction was inhibited in the presence of an excess of Zn 2ϩ , which suggests that this peptidase belongs to the M1 family of zinc-binding metalloproteases. Aminopeptidase II from S. cerevisiae is believed to be involved in the cellular supply of leucine from externally offered leucine-containing di-or tripeptide substrates (30). We found similar properties of S. boulardii peptidase against the peptide L-Leu-Gly-Gly.…”

“…Zinc-binding metalloproteases can be grouped together as a superfamily known as the metzincins on the basis of their sequence similarity (27). Family M1 includes bacterial aminopeptidase N (EC 3.4.11.2), mammalian aminopeptidase N (EC 3.4.11.2), mammalian glutamyl aminopeptidase (EC 3.4.11.7) or aminopeptidase A, leukotriene A-4 hydrolase (EC 3.3.2.6) (28), alanine/arginine aminopeptidase from S. cerevisiae (gene AAP1) (29), and aminopeptidase II (gene APE2) from S. cerevisiae (30). The similarities between the sequences of human, rat, and yeast aminopeptidases that contain the (29,30).…”

Aminopeptidase C of Aspergillus niger Is a Novel Phenylalanine Aminopeptidase

“…F-pNA was used as a substrate for the determination of the pH optimum, the pH stability, and the temperature stability of the purified enzyme. The optimal pH is 5, which is different from the pH optima reported for A. niger ApsA and other intracellular aminopeptidases, such as the yeast aminopeptidases which all have pH optima around 7.5 (1,4,7,11).…”

Cloning and Characterization of a Novel Insulin-regulated Membrane Aminopeptidase from Glut4 Vesicles

“…vp165 as an Aminopeptidase-Comparison of the predicted amino acid sequence of vp165 with those in the sequence data banks revealed that the large 785-amino acid C-terminal portion of vp165 contained regions with high homology to domains conserved between the mammalian aminopeptidases A and N (23, 24), aminopeptidase yscII from Saccharomyces cerevisiae (25), and aminopeptidase N from Lactococcus lactis (26), as well as the recently described thyrotropin-releasing hormone degrading enzyme from rat (27). The similarity is highest in a stretch of 349 amino acids, where the overall identity to vp165 ranges between 36% for thyrotropin releasing hormone degrading enzyme and 45% for aminopeptidase yscII (Fig.…”