[28] Cleavage at aspartic acid

Schultz, Julius

doi:10.1016/s0076-6879(67)11030-6

Cited by 130 publications

(78 citation statements)

References 3 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However. there is a wealth of information regarding the acid susceptibility of specific peptide bonds (Light, 1967;Schultz, 1967;Zhou & Smith, 1990), particularly those that can undergo acyltransfer reactions through either an ester or anhydride intermediate. Perhaps the best known is the Asp-Pro bond, which is cleaved under mildly acidic conditions when other peptide bonds are largely unaffected.…”

Section: Lshmentioning

confidence: 99%

See 1 more Smart Citation

A biomimetic strategy in the synthesis and fragmentation of cyclic protein

1998

View full text Add to dashboard Cite

This paper describes a simple biomimetic strategy to prepare small cyclic proteins containing multiple disulfide bonds. Our strategy involves intramolecular acyl transfer reactions to assist both the synthesis and fragmentation of these highly constrained cyclic structures in aqueous solution. To illustrate our strategy, we synthesized the naturally occurring circulin B and cyclopsychotride (CPT), both consisting of 3 1 amino acid residues tightly packed in a cystine-knot motif with three disulfide bonds and an end-to-end cyclic form. The synthesis of these small cyclic proteins can be achieved by orthogonal ligation of free peptide thioester via the thia zip reaction, which involves a series of reversible thiolthiolactone exchanges to arrive at an a-amino thiolactone, which then undergoes an irreversible, spontaneous ring contraction through an S,N-acyl migration to form the cyclic protein. A two-step disulfide formation strategy is employed for obtaining the desired disulfide-paired products. Partial acid hydrolysis through intramolecular acyl transfer of X-Ser, X-Thr, Asp-X, and Glu-X sequences is used to obtain the assignment of the circulins disulfide bond connectives. Both synthetic circulin B and CPT are identical to the natural products and, thus, the total synthesis confirms the disulfide connectivity of circulin B and CPT contain a cystine-knot motif of 1-4,2-5, and 3-6. In general, our strategy, based on the convergence of chemical proteolysis and aminolysis of peptide bonds through acyl transfer, is biomimetic and provides a useful approach for the synthesis and characterization of large end-to-end cyclic peptides and small proteins.

show abstract

Section: Lshmentioning

confidence: 99%

“…More than 30 years ago, several laboratories had already firmly established that partial acid hydrolysis is not a random process in the cleavage of peptide bonds (Light, 1967;Schultz, 1967). However, problem of identifying overlapping peptides proved to be too laborious for practical use.…”

Section: Partial Acid Hydrolysis For Assignment Of Disuljide Bondsmentioning

confidence: 99%

A biomimetic strategy in the synthesis and fragmentation of cyclic protein

1998

View full text Add to dashboard Cite

show abstract

“…At present, the appearance of this component is inexplicable. Schultz has stated that cleavage of protein at peptide bonds linking aspartic acid or asparagine occurs in acid solutions [31].…”

Section: Amino Acid Compositionmentioning

confidence: 99%

Subunit Structure and Multifunctional Properties of Yeast Phosphoglyceromutase

Sasaki

Utsumi

Sugimoto

et al. 1976

European Journal of Biochemistry

View full text Add to dashboard Cite

A new and efficient method for preparation of pure phosphoglyceromutase from baker's yeast (Saccharomyces cerevisiae) is described. Proteolytic alterations of the enzyme during extraction can be minimized by grinding the dried yeast with aluminium oxide at low temperature. Yeast phosphoglyceromutase contains four highly similar, probably identical subunits of molecular weight 28000, a conclusion based on the following observations. Polyacrylamide gel electrophoresis containing dodecylsulphate or urea gives a single band, indicating that the enzyme is composed of four subunits similar in their molecular weight and net charge. Cyanogen bromide cleavage and, tryptic digestion of the enzyme yield the number of peptides expected for identical subunits from the amino acid composition analysis. The purred phosphoglyceromutase preparation has bisphosphoglyceromutase activity synthesizing‐2,3‐bisphosphoglycerate from 1,3‐bisphosphoglycerate and 3‐phosphoglycerate. It has been reported that yeast phosphoglyceromutase catalyzes the hydrolysis of 2,3‐bisphosphoglycerate at the same active site which catalyzes the phosphoglyceromutase reaction [Sasaki, R. et al. (1971) Biochim. Biophys. Acta, 227, 584–594, 595–607]. Immunological studies and chemical modification experiments indicate that bisphosphoglyceromutase activity also is due to the phosphoglyceromutase protein and involves amino groups which have been shown to be essential for the other two activities.

show abstract

“…The PNA sequence was determined using two less common methods, limited acid cleavage at Asp-Pro peptide bonds [19] and stronger acid cleavage at Asp residues [20]. Both gave excellent results and deserve to be used more widely.…”

Section: Discussionmentioning

confidence: 99%

The amino acid sequence of peanut agglutinin

Young

Johnston

Watson

1991

European Journal of Biochemistry

View full text Add to dashboard Cite

The amino acid sequence of peanut (Arachis hypogaea) agglutinin was determined from three major fragments obtained by mild acid cleavage at Asp-Pro peptide bonds. The sequence of 236 amino acids has residues identical to those that form the metal-binding site and the hydrophobic pocket in concanavalin A and other lectins, although the overall similarity is only 42%. In the segments of peanut agglutinin that correspond to the four loops that form the carbohydrate-binding site in concanavalin A and favin, several central residues are homologous, while others show changes to smaller side chains, such as Tyr --f Gly. The carbohydrate-binding site of peanut agglutinin may therefore have a similar peptide-backbone architecture, but form a considerably more open cleft.Peanut agglutinin (PNA) is a legume seed lectin that is specific for D-galactose residues at non-reducing terminal positions of glycoconjugates [l]. This specificity has made it useful for histochemical and cell-surface studies. The lectin recognises both a-and P-galactosides, and the disaccharide of highest known affinity is the T-antigen structure Gal(b1-3)GalNAc [2] (GalNAc, N-acetyl-D-galactosamine). Detailed studies of the interactions of PNA with monosaccharides and disaccharides have been made by UV difference [3,4], fluorescence [5, 61 and NMR [7, 81 spectroscopy, and a model of the binding process has been proposed [8]. PNA is therefore one of the best characterized lectins in its functional aspects, but far less is known about its structure. It is a tetramer of subunits of molecular mass 27 kDa [9], each with one bindingsite [3], and is not a glycoprotein [I]. PNA has been successfully crystallized for X-ray studies in two laboratories [lo, 1 I].A partial amino acid sequence was reported by Lauwereys et al. [I21 who encountered difficulties with multiple genetic forms and poorly soluble peptides. The former are to be expected in view of the multiple bands seen in isoelectric focussing and chromatofocussing experiments [13,14]. We have reinvestigated the amino acid sequence of PNA and report a complete sequence together with data on the genetic variation at some positions. The sequences of the peptide loops in the putative carbohydrate-binding-site region are discussed in relation to the X-ray results for the D-mannose-specific lectins concanavalin A Abbreviations. PNA, peanut agglutinin; GalNAc, N-acetyl-Dgalactosamine. MATERIALS AND METHODS Purqication of the proteinPeanut agglutinin was purified from buffer extracts of delipidated peanut meal by affinity chromatography on a lactose-substituted agarose matrix [3]. The initial PNA sample was material used for spectroscopic studies [3,4,7, 81; subsequent samples were prepared identically from the same batch of peanuts. Chemical cleavageThe protein was cleaved at Asp-Pro peptide bonds by limited acid hydrolysis [19]. PNA (22 mg) was dissolved in 2.28 ml 88% formic acid, 1.72 ml water was added and the solution was incubated at 37°C for 96 h. The digest was diluted and freeze-dried. The fragments...

show abstract

[28] Cleavage at aspartic acid

Cited by 130 publications

References 3 publications

A biomimetic strategy in the synthesis and fragmentation of cyclic protein

A biomimetic strategy in the synthesis and fragmentation of cyclic protein

Subunit Structure and Multifunctional Properties of Yeast Phosphoglyceromutase

The amino acid sequence of peanut agglutinin

Contact Info

Product

Resources

About