A determination of the positions of disulphide bonds in Paim I, α-amylase inhibitor fromStreptomyces corchorushii, using fast atom bombardment mass spectrometry

Akashi, Satoko; Hirayama, Kazuo; Seino, Tadatoshi; Ozawa, Shinobu; Fukuhara, Ken-ichi; Oouchi, Naoki; Murai, Asao; Arai, Motoo; Murao, Sawao; Tanaka, Kazuo; Nojima, Ittetsu

doi:10.1002/bms.1200151006

Cited by 24 publications

(5 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The reactive site contains the triad Trp18, Arg19, and Tyr20 and tightly binds in an equimolar complex with α-amylase. These amino acids are located on the β-loop on the surface of the proteins so that the hydrophobic part of the arginine residue is held between two stacked aromatic side-chains with the guanidinium group exposed to water [89][90][91][92][93].…”

Section: Inhibitors From Streptomycesmentioning

confidence: 99%

Natural Inhibitors of Mammalian α-Amylases as Promising Drugs for the Treatment of Metabolic Diseases

Kalinovskii,

Sintsova,

Gladkikh

et al. 2023

IJMS

View full text Add to dashboard Cite

α-Amylase is a generally acknowledged molecular target of a distinct class of antidiabetic drugs named α-glucosidase inhibitors. This class of medications is scarce and rather underutilized, and treatment with current commercial drugs is accompanied by unpleasant adverse effects. However, mammalian α-amylase inhibitors are abundant in nature and form an extensive pool of high-affinity ligands that are available for drug discovery. Individual compounds and natural extracts and preparations are promising therapeutic agents for conditions associated with impaired starch metabolism, e.g., diabetes mellitus, obesity, and other metabolic disorders. This review focuses on the structural diversity and action mechanisms of active natural products with inhibitory activity toward mammalian α-amylases, and emphasizes proteinaceous inhibitors as more effective compounds with significant potential for clinical use.

show abstract

Section: Inhibitors From Streptomycesmentioning

confidence: 99%

Natural Inhibitors of Mammalian α-Amylases as Promising Drugs for the Treatment of Metabolic Diseases

Kalinovskii,

Sintsova,

Gladkikh

et al. 2023

IJMS

View full text Add to dashboard Cite

show abstract

“…The specific algorithms described in this paper have been designed for the use of integer variables such as the nominal FAB-MS MH + data, since, up to now, these are the most used data for the assignment of protein disulphide bridges [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16]. However, the average mass data usually provided by conventional instruments equipped for electrospray ionisation (ESI) or matrix-assisted laser desorption/ionisation (MALDI) can also be processed by the method.…”

Section: Required Datamentioning

confidence: 99%

“…both containing two cysteine residues, can be linked by either one or two S-S bridges, giving rise to structures with different mass values. This aspect should be considered even if the native protein does not contain free thiol groups, as the reduction of S-S bridges can take place under FAB conditions [4,13,17]. The algorithm has been designed to consider even the presence of either homoserine or the corresponding lactone at the C-terminus of peptides derived from CNBr hydrolysis of the native protein.…”

Section: Searching For the Disulphide Bridgesmentioning

confidence: 99%

Assignment of protein disulphides by a computer method using mass spectrometric data

Caporale¹,

Sepe²,

Caruso³

et al. 1996

FEBS Letters

View full text Add to dashboard Cite

We designed a computer program for the assignment of protein disulphides using mass spectrometric data. All the theoretical linear peptides containing from one to three cysteines are generated on the basis of the protein sequence. Their combination ways are determined in order to create all the possible disulphide-bridged fragments containing from two to six cysteines and to calculate their molecular weight. One, two and three S-S bridges per linked fragment were considered, taking into account the possibility that the fragments contain unabridged residues. The mass data obtained from the spectral analysis of peptide mixtures of the digested protein are then associated to the fitting structures of disulphide-bridged peptides, giving rise to the primary output. This output can then be screened by using information on the specificity of the proteolytic agent(s) used and/or any further mass data provided by Edman degradation and/or carboxypeptidase treatment of the peptide mixtures. The need for such a computer aid is discussed and examples of application are given.Key words: Disulfide bridge; Protein structure; Computer method; Mass spectrometry and the number of fragments, a single mass signal can be associated to many clusters of bridged peptides, with the ambiguity remaining still unsolved even after one step of chemical and/or enzymatic degradation [13].This paper describes the design and application of a computer program analyzing the possible solutions and filtering the results on the basis of new data obtained after degradation and/or information on the specificity of the proteolytic agent(s). We also suggest that such a computer aid can be indispensable in many cases for the unambiguous assignment of disulphide bridges. Materials and methodsThe mass data and experimental procedures of the two proteins used in the application examples have already been reported [13,15]. Programs were written using Microsoft QuickBASIC (version 1.00b) and implemented on an Apple Macintosh LC 475 computer. The operative system was System 7.1. The compiled applications are compatible with all Apple Macintosh computers. Results and discussion

show abstract

“…In recent years, mass spectrometric techniques have emerged as a major tool for disulphide bond determination and localisation [11][12][13][14]. This paper is concerned with the detection and assignment of disulphide bonds in the 1-30 synthetic peptide by means of ESI-MS in combination with chemical and enzymatic cleavage and reversedphase high-performance liquid chromatography (RP-HPLC).…”

Section: Introductionmentioning

confidence: 99%

Detection and localisation of disulphide bonds in a synthetic peptide reproducing the sequence 1-30 ofPar j1.0101 by electrospray ionisation mass spectrometry

et al. 2001

View full text Add to dashboard Cite

The structural characterisation of a synthetic peptide reproducing the sequence 1-30 of Par j 1.0101, a major allergenic protein present in the pollen of Parietaria judaica, by combined use of chemical and enzymatic cleavage, reversed-phase high-performance liquid chromatography and electrospray ionisation mass spectrometry (ESI-MS), is described. Direct ESI-MS of the synthetic peptide after reaction with methyl iodide showed that the product is a mixture of two peptides: one form in which two out of the four cysteine residues present in the sequence are oxidised and a minor amount of another form in which all the cysteines are fully reduced. It was ascertained, using the combined procedure indicated above and without prior separation of the two species, that the disulphide bond in the partially oxidised form is located between cysteines 29 and 30. These results show the usefulness of this approach for characterising synthetic peptides containing multiple cysteine residues in the sequence.

show abstract

A determination of the positions of disulphide bonds in Paim I, α-amylase inhibitor fromStreptomyces corchorushii, using fast atom bombardment mass spectrometry

Cited by 24 publications

References 23 publications

Natural Inhibitors of Mammalian α-Amylases as Promising Drugs for the Treatment of Metabolic Diseases

Natural Inhibitors of Mammalian α-Amylases as Promising Drugs for the Treatment of Metabolic Diseases

Assignment of protein disulphides by a computer method using mass spectrometric data

Detection and localisation of disulphide bonds in a synthetic peptide reproducing the sequence 1-30 ofPar j1.0101 by electrospray ionisation mass spectrometry

Contact Info

Product

Resources

About