Adsorbents for affinity chromatography. Use of N-hydroxysuccinimide esters of agarose

Cuatrecasas, Pedro; Parikh, Indu

doi:10.1021/bi00762a013

Cited by 452 publications

(147 citation statements)

References 17 publications

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“…The modification of Tyr 4 is shown by the absence of the unmodified (2-5) internal ion, and the presence of the (2-5)ϩAc internal ion (Figure 1e). Thus, it is apparent that the succinimidyl esters are selective for the N-terminus over tyrosine, but do react with tyrosine, as was observed in a previous study [12][13][14]. In these same studies, it was also shown that histidine could be transiently modified by the succinimidyl esters [12,13].…”

Section: Reactivity Of N-hydroxysuccinimidyl Acetatesupporting

confidence: 62%

“…However, in the early bio-chemical characterization of these active esters as cross-linking reagents, it was found that they showed varying reactivity with the following residues: histidine imidazole Ͼ primary amino groups ӷ cysteine thiolate Ϸ tyrosine phenolate Ͼ serine [12][13][14]. Therefore, it is possible that these reagents can be used to cross-link amino acids other than the N-terminus and lysine.…”

Section: Reactivity Of N-hydroxysuccinimidyl Acetatementioning

confidence: 99%

“…This is especially true if the specificity of the reagent can be adjusted based on the reaction conditions. In considering the possible cross-links formed by the succinimidyl esters, the following three points must be considered: it was found previously that the N-acylimidazole product (formed when His is modified) is transient and hydrolyses quickly, cystine is commonly unreactive as it is often involved in disulfide bonds, and the kinetics of serine reactivity are quite slow and cannot compete with the other fast reactions (e.g., with N-terminus, lysine, and tyrosine) [12][13][14][15]. Therefore, the primary amino group-containing amino acids and tyrosine are the most likely amino acids to form stable cross-links with these esters in soluble proteins.…”

Section: Reactivity Of N-hydroxysuccinimidyl Acetatementioning

confidence: 99%

“…Using angiotensin I and oxidized insulin-␤-chain we show that the succinimidyl ester crosslinkers can be used to form cross-links not only between lysines, but also between lysine and tyrosine and even between tyrosines. In the early development and characterization of these cross-linkers [12][13][14], the reactivity towards tyrosine and other amino acids (e.g., histidine, cystine, and serine) was noted, but has largely been overlooked in recent studies. The reactivity of histidine and serine can generally be ignored owing to the instability of the N-acylimidazole product (histidine) and the slow reactivity of serine.…”

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confidence: 99%

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Strategy for selective chemical cross-linking of tyrosine and lysine residues

Leavell

Novák

Behrens

et al. 2004

J. Am. Soc. Mass Spectrom.

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Chemical cross-linking of proteins combined with mass spectral analysis is a powerful technique that can be utilized to yield protein structural information, such as the spatial arrangement of multi-protein complexes or the folding of monomeric proteins. The succinimidyl ester cross-linking reagents are commonly used to cross-link primary amine-containing amino acids (N-terminus and lysine). However, in this study they were used to react with tyrosines as well, which allowed for the formation of cross-links between two primary amines, one primary amine and one tyrosine, or two tyrosines. This result is extremely important to the chemical cross-linking community for two reasons: (1) all possible cross-linked residues must be considered when analyzing data from these experiments to generate correct distance constraints and structural information, and (2) utilizing the versatility of these cross-linking reagents allows more information content to be generated from a single cross-linking reagent, which may increase the number of cross-links obtained in the experiment. Herein, we study the reactivity of the succinimidyl ester labeling and cross-linking reagents with angiotensin I and oxidized insulin ␤-chain. Using the succinimidyl acetate labeling reagent, the reactivity of the N-terminus was found to be greater than either lysine or tyrosine. However, a selectivity of the cross-linking reagent was observed for either tyrosine or lysine depending on the pH of the reaction solution. In acidic pH, it was observed that tyrosine was more reactive, while in alkaline pH lysine was more reactive. Exploiting this selectivity predominantly N-terminustyrosine or tyrosine-tyrosine cross-links were favored at acidic pH, while N-terminus-tyrosine or tyrosine-lysine cross-links were favored at alkaline pH. I nter-molecular chemical cross-linking has a long history as a tool for the study of the quaternary structure of protein complexes [1,2], and many years ago it was suggested that intra-molecular cross-linking could be used as a method of obtaining distance constraints that would be useful in developing structural models of proteins [3]. However, the promise of intra-molecular crosslinking for structural modeling has only recently been enabled by state of the art mass spectrometric methods that make determination of the cross-linked residues in a protein practical on a reasonable time scale and with small quantities of protein. The first study to derive a sufficient number of intra-molecular distance constraints to develop a structural model for a protein used fibroblast growth factor 2 (FGF-2) as a test case [4]. FGF-2 is a 17 kDa protein, which has 14 lysines evenly dispersed in its 155 residue sequence. Using only the commercially available primary amine reactive cross-linker bis(sulfosuccinimidyl) suberate, 18 cross-links were found in FGF-2 in this study, of which 15 provided useful distance constraints for determining the fold family of the protein. Using these 15 constraints it was possible to assign the FGF2 to the corr...

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Section: Reactivity Of N-hydroxysuccinimidyl Acetatesupporting

confidence: 62%

Section: Reactivity Of N-hydroxysuccinimidyl Acetatementioning

confidence: 99%

Section: Reactivity Of N-hydroxysuccinimidyl Acetatementioning

confidence: 99%

mentioning

confidence: 99%

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Strategy for selective chemical cross-linking of tyrosine and lysine residues

Leavell

Novák

Behrens

et al. 2004

J. Am. Soc. Mass Spectrom.

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“…It could thus be expected that more vigorous coupling conditions, favoring a higher degree of substitution, would still leave the enzyme with its dehydrogenase function intact. It was also found that rather prolonged reaction times combined with the presence of N-hydroxysuccinimide [24] promoted coupling without seriously affecting the specific activity. The coupling procedure ( Fig.…”

Section: Coupling Of N a D Analogue To The Enzymementioning

confidence: 98%

Covalent Binding of an NAD Analogue to Liver Alcohol Dehydrogenase Resulting in an Enzyme‐Coenzyme Complex not Requiring Exogenous Coenzyme for Activity

Månsson

Larsson

Mosbach

1978

European Journal of Biochemistry

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The NAD analogue, N6‐[N‐(6‐aminohexyl)carbamoylmethyl]‐NAD, was covalently bound to horse liver alcohol dehydrogenase in a carbodiimide‐mediated reaction and in such a way that it was active with the very same enzyme molecule to which it was coupled. The degree of substitution, i.e. the number of NAD analogues per enzyme subunit, could be varied (0.3–1.6). In one preparation 1.6 coenzyme molecules were bound per subunit; the alcohol dehydrogenase activity of this preparation was 40% of the activity obtained after addition of free NAD in excess. It was calculated that every fourth active site of this preparation was provided with a covalently bound functioning coenzyme analogue, and that this analogue had a cycling rate of about 40000 cycles/h in a coupled substrate assay. The presence of the covalently bound coenzyme made the active sites difficult to inhibit with a competitive inhibitor. For example, 10 mM AMP inhibited the activity of the preparation by 50% whereas a reference system containing native alcohol dehydrogenase was inhibited by 80% in spite of the fact that the reference system contained about 20000 times as high a concentration of coenzyme.

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Different synthetic routes to obtain hydrophilic matrices

Álvarez

Strumia

2000

J. Polym. Sci. A Polym. Chem.

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Adsorbents for affinity chromatography. Use of N-hydroxysuccinimide esters of agarose

Cited by 452 publications

References 17 publications

Strategy for selective chemical cross-linking of tyrosine and lysine residues

Strategy for selective chemical cross-linking of tyrosine and lysine residues

Covalent Binding of an NAD Analogue to Liver Alcohol Dehydrogenase Resulting in an Enzyme‐Coenzyme Complex not Requiring Exogenous Coenzyme for Activity

Different synthetic routes to obtain hydrophilic matrices

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