A new acrylamido buffer has been synthesized, for use in isoelectric focusing in immobolized pH gradients. This compound (2-acrylamido glycolic acid) has a pK = 3.1 (at 25°C, 20 mM concentration during titration) and is used, by titration with the pK 9.3 Immobiline, to produce a linear pH gradient in the pH 2.5-3.5 interval. Pepsin (from pig stomach) focused in this acid pH gradient is resolved into four components, two major (with pI values 2.76 and 2.78) and two minor (having pI values 2.89 and 2.90). This is the first time that such strongly acidic proteins could be focused in an immobilized pH gradient. Even in conventional isoelectric focusing in amphoteric buffers it has been impossible to focus reproducibly very-low-pI macromolecules.

Focusing of pepsin in strongly acidic immobilized pH gradients / P.G.Righetti, M. Chiari, P.K. Sinha, E. Santaniello. - In: JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS. - ISSN 0165-022X. - 16:2-3(1988), pp. 185-192.

Focusing of pepsin in strongly acidic immobilized pH gradients

E. Santaniello
Ultimo
1988

Abstract

A new acrylamido buffer has been synthesized, for use in isoelectric focusing in immobolized pH gradients. This compound (2-acrylamido glycolic acid) has a pK = 3.1 (at 25°C, 20 mM concentration during titration) and is used, by titration with the pK 9.3 Immobiline, to produce a linear pH gradient in the pH 2.5-3.5 interval. Pepsin (from pig stomach) focused in this acid pH gradient is resolved into four components, two major (with pI values 2.76 and 2.78) and two minor (having pI values 2.89 and 2.90). This is the first time that such strongly acidic proteins could be focused in an immobilized pH gradient. Even in conventional isoelectric focusing in amphoteric buffers it has been impossible to focus reproducibly very-low-pI macromolecules.
Settore BIO/10 - Biochimica
Article (author)
File in questo prodotto:
Non ci sono file associati a questo prodotto.
Pubblicazioni consigliate

Caricamento pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/195738
Citazioni
  • ???jsp.display-item.citation.pmc??? 0
  • Scopus 29
  • ???jsp.display-item.citation.isi??? 22
social impact