Size restriction on utilization of peptides by amino acid auxotrophs of Neurospora crassa

Wolfinbarger, Lloyd; Marzluf, George A.

doi:10.1128/jb.122.3.949-956.1975

Cited by 11 publications

(12 citation statements)

References 12 publications

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“…These workers concluded that, at present, it is impossible to decide whether the size limit occurs at the level of the lipopolysaccharide or of the peptidoglycan. In Neurospora crassa, mutants containing porous cell walls were used to show that the peptide transport system itself is incapable of handling peptides that, on the average, contain more than five residues (21). In the present study, we found that all tetrapeptides containing leucine or lysine failed to serve the nutritional requirements of strain Z1-2D.…”

Section: Discussionmentioning

confidence: 49%

“…Previous studies with microorganisms concluded that size limits to peptide uptake could result from either a sieving action of the cell envelope (1,15) or from the failure of the peptide transport system to bind peptides above a given size (21). The size restriction to peptide transport in E. coli has been discussed recently in detail by Barak and Gilvarg (1).…”

Section: Discussionmentioning

confidence: 99%

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Peptide transport in yeast: utilization of leucine- and lysine-containing peptides by Saccharomyces cerevisiae

Marder¹,

Becker²,

Naider³

1977

J Bacteriol

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A variety of leucine-containing diand tripeptides and two lysine-containing dipeptides supported the growth of strain Z1-2D, a leucine, lysine auxotroph of Saccharomyces cerevisiae. However, (Lys)2, (Lys)3, (Lys)4, and (Lys)5 as well as Gly-Leu-Gly, three tetra-and one pentapeptide containing leucine were not utilized by the mutant. Cellular peptidases released leucine or lysine from all of these non-growth-supporting peptides, suggesting that the failure of strain Zl-2D to utilize these compounds reflects their failure to enter the yeast. Competition studies employing phenylalanine or non-leucine-containing peptides showed that the uptake of peptides into S. cerevisiae Z1-2D is distinct from that of amino acids and that diand oligopeptides may share a common transport system. The failure of strain Z1-2D to utilize any peptide larger than (Leu)3 may indicate a transport size limit. Such a size limit would influence the construction of models that explain the action of yeast mating factors.The transport and utilization of peptides have been investigated extensively in Escherichia coli (1,7,14,16) and the mammalian gut (1,7,11,16). The only previously published reports of peptide transport in Saccharomyces cerevisiae deal with the uptake of methioninecontaining peptides by a strain G1333, a methionine auxotroph (4, 12). Information concerning the effect of such factors as amino acid sequence, derivatization of end groups, and peptide length on peptide utilization was determined by using strain G1333. Preliminary studies with this organism, however, left unanswered many important questions concerning the structural requirements for peptide transport in S. cerevisiae. To determine whether peptides comprised entirely of residues other than methionine could also enter S. cerevisiae and to learn more about the structural requirements for peptide transport in this organism, we examined peptide utilization in another auxotroph of this eucaryote.This report deals with transport in S. cerevisiae Z1-2D, a strain that requires both lysine and leucine. In a parallel investigation, we have followed the uptake of radioactively labeled (Met)3 by a wild-type strain of S. cerevisiae (3). MATERIALS AND METHODSYeast strain and growth conditions. Strain Zi-2D, a lys leu auxotroph of S. cerevisiae, was re-ceived from Nasim Khan, Brooklyn College, City University of New York. This haploid strain was derived from a cross between strains KC-372 (X. Mortimer, University of California, Berkeley) and 1323-1B (D. C. Hawthorne, University of Washington, Seattle). The minimal growth medium used for all growth experiments was yeast nitrogen base without amino acids (Difco Laboratories, Detroit, Mich.). This medium was prepared in a ten-timesconcentrated form (6.7 g of yeast nitrogen base in 100 ml of water), 5 g of dextrose was added, and the solution was filter sterilized (Nalge, 0.22 ,um) and refrigerated. All amino acids and peptides were filter sterilized.The strain was maintained on slants composed of 1% yeast extract, 2% peptone, 2% dext...

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Section: Discussionmentioning

confidence: 49%

Section: Discussionmentioning

confidence: 99%

Peptide transport in yeast: utilization of leucine- and lysine-containing peptides by Saccharomyces cerevisiae

Marder¹,

Becker²,

Naider³

1977

J Bacteriol

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“…We previously reported that peptides with a mean number of amino acid residues greater than five would not support growth of a leucine auxotroph (leu-2) of Neurospora crassa because this organism was unable to transport larger peptides (10). The exception to this observation occurred when a minimal amount of growth was permitted, which presumably resulted in the production of extracellular peptidohydrolytic activity with subsequent digestion of the oligopeptides to transportable amino acids and small (three to five residue) peptides (10).…”

mentioning

confidence: 99%

“…The peptides to be used as inducers in this study were obtained as previously described (10) by passage of a partial enzymatic digest of peptone, Neopeptone (Difco Laboratories), over a Sephadex G-15 column (100 by 2.5 cm) equilibrated with sodium phosphate buffer (0.01 M, pH 7.0) that also contained 0.1 M sodium chloride. The eluted peptides were collected as fractions, and their concentrations (1 to 2 mM) were determined with trinitrobenzene sulfonic acid (10). The mean number of amino acid residues in peptides eluting in a given fraction was determined by the ratio of absorbance (trinitrobenzene sulfonic acid assay) before and after acid hydrolysis (10).…”

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confidence: 99%

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Peptide utilization by nitrogen-starved Neurospora crassa

Wolfinbarger¹,

Snyder²,

Castellano³

1983

J Bacteriol

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Peptides ranging in size from a mean number of 30 residues down to dipeptides supported growth of a leucine auxotroph when used as both a nitrogen and leucine source. Under nitrogen-limiting conditions, the peptides induced extracellular peptidohydrolytic activity, hydrolyzing peptides to monomer amino acids. Growth of a leu-2 mutant of Neurospora crassa on those peptides transportable by the oligopeptide transport system did not result in induction of hydrolytic activity, whereas growth of a leu-2; gltR mutant on these same peptides resulted in induction of peptidohydrolytic activity. The induced extracellular proteolytic activity was shown to be analogous to that inducible by growth on proteins, e.g., bovine serum albumin.

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Phase I study of beta-alanyl-melphalan as a potent anticancer drug

Tsay

Wolfinbarger

1987

Cancer Chemother. Pharmacol.

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The dipeptide beta-alanyl-melphalan was synthesized and tested for its potential anticancer activity. It is shown to possess considerable toxicity toward Ehrlich ascites tumor cells and 3T3 mouse embryo cells in in vitro toxicity assays and is a potent anticancer agent when used in vivo in traditional phase I chemotherapy assays. The potential role for small peptide transport mechanisms in transportation of anticancer agents is discussed.

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Size restriction on utilization of peptides by amino acid auxotrophs of Neurospora crassa

Cited by 11 publications

References 12 publications

Peptide transport in yeast: utilization of leucine- and lysine-containing peptides by Saccharomyces cerevisiae

Peptide transport in yeast: utilization of leucine- and lysine-containing peptides by Saccharomyces cerevisiae

Peptide utilization by nitrogen-starved Neurospora crassa

Phase I study of beta-alanyl-melphalan as a potent anticancer drug

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