Affinity chromatography of plasma proteins (guanidinobenzoatase): use of mimetic matrices and mimetic soluble ligands to prevent the binding of albumin on target affinity matrices

Murza, Adrian; Aguilar, Alfredo Robledo; Fernández‐Lafuente, Roberto; Guisán, José M.

doi:10.1016/s0378-4347(99)00279-0

Cited by 6 publications

(4 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…GB Purification. GB for in vitro studies was extracted from the ascitic fluid of a mouse Ehrlich tumor, and purified by using an agmatine-CH Sepharose 4B affinity column as described in reference (18). In the last work, a double purification strategy based on the use of mimetic matrixes as well as soluble mimetic ligands is described.…”

Section: Methodsmentioning

confidence: 99%

“…Since the structure of the active site of GB has not been yet reported, the interaction of 9AA with other serine-active enzymes such as penicillin G-acylase (PGA), trypsin, and R-chymotrypsin, which display a similar catalytic activity, has been also investigated to afford more information about the GB active site structure and the 9AA-GB photodynamic mechanism. Moreover, the interaction with bovine serum albumin (BSA) was also checked as this carrier protein interferes in the purification of GB from the biological fluids employed as GB sources (18).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Interaction of the Antitumor Drug 9-Aminoacridine with Guanidinobenzoatase Studied by Spectroscopic Methods: A Possible Tumor Marker Probe Based on the Fluorescence Exciplex Emission

et al. 2000

Self Cite

View full text Add to dashboard Cite

Fluorescence spectroscopy, surface-enhanced Raman spectroscopy (SERS), and analytical centrifugation are applied in this work to study the interaction of the antitumor drug 9-aminoacridine (9AA) with a trypsin-like protease, guanidinobenzoatase (GB), extracted from an Erlich tumor. As a consequence of this interaction, a strong 9AA exciplex emission can be detected at a certain drug and enzyme concentration. The 9AA exciplex emission was also studied for 9AA interacting with others serin proteases: alpha-chymotrypsin, trypsin, and penicillin G-acylase (PGA), as well as with bovine serum albumin (BSA) in order to obtain information about the active center of GB. We have found that the exciplex 9AA emission may be induced by a ring-stacking interaction between the monomeric drug, under the amino form, and an aromatic residue placed in the catalytic site of the protein. The results derived from Raman spectroscopy corroborate this interaction mechanism, as demonstrated by the existence of typical protonated amino 9AA marker bands as well as an important modification of the ring vibrations, thus indicating the existence of an interaction through ring stacking. The analytical centrifugation technique was applied to study the GB association in aqueous solution, demonstrating that the 9AA/GB interaction depends on the enzyme quaternary structure. An interaction of 9AA with an associate form of GB, which may be the actual enzyme active form, is suggested.

show abstract

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Interaction of the Antitumor Drug 9-Aminoacridine with Guanidinobenzoatase Studied by Spectroscopic Methods: A Possible Tumor Marker Probe Based on the Fluorescence Exciplex Emission

et al. 2000

Self Cite

View full text Add to dashboard Cite

show abstract

“…The GB for the in vitro studies was purified from a mouse Ehrlich tumor, which was extracted in a single step by using an agmatine‐CH Sepharose 4B affinity column as described elsewhere 26. GB elution was carried out in the presence of the soluble substrate NPGB.…”

Section: Methodsmentioning

confidence: 99%

Surface‐enhanced Raman and steady fluorescence study of interaction between antitumoral drug 9‐aminoacridine and trypsin‐like protease related to metastasis processes, guanidinobenzoatase

2001

Self Cite

View full text Add to dashboard Cite

Fluorescence spectroscopy and surface-enhanced Raman spectroscopy (SERS) were applied to study the interaction of the antitumoral drug 9-aminoacridine (9AA) with a trypsin-like protease guanidinobenzoatase (GB) extracted from a mouse Erlich tumor. As a consequence of this interaction, a strong 9AA exciplex emission was detected in the emission fluorescence spectra at certain drug and enzyme concentrations. A SERS study was accomplished on silver colloids at several excitation wavelengths in order to obtain more information about the interaction mechanism. The results derived from Raman spectroscopy indicated that 9AA in the amino monomeric form may interact with the enzyme by means of two different bonds: an ionic bond with a negatively charged amino acid and a ring stacking interaction with an aromatic residue placed in the catalytic site of GB. This interaction mechanism was responsible for a strong exciplex emission detected at a longer wavelength than the expected value of the normal fluorescence emission. Moreover, the GB concentration dependence of the interaction suggested that the drug was sensitive to the quaternary structure of the enzyme.

show abstract

“…Thus, if many ligands are present on the enzyme support, they may be able to interact by multiple points with any biomacromolecule (Fuentes et al, 2007, Murza et al, 1999.…”

Section: Coupled Immobilization/purification Of Enzymes and Proteins mentioning

confidence: 99%

Strategies for the one-step immobilization–purification of enzymes as industrial biocatalysts

Barbosa

Ortíz

Berenguer-Murcia

et al. 2015

Biotechnology Advances

Self Cite

600

318

View full text Add to dashboard Cite

Enzyme immobilization and purification are two steps that are usually required in the development of any industrial biocatalyst. In this review, we detail the efforts performed to couple the purification and the immobilization of industrial enzymes in a single step. The use of antibodies (versus the target enzyme or versus some domains), the development of specific domains with affinity for some specific supports or just to increase the affinity for standard ones (ionic exchangers, silicates) will be revised. We will show how the control of the immobilization conditions may convert some unspecific supports in largely specific ones. The development of tailormade heterofunctional supports as a tool to immobilize-stabilize-purify some proteins will be discussed in deep, using low concentration of adsorbents groups and a dense layer of groups able to give an intense multipoint covalent attachment. The final coupling of mutagenesis and tailor made supports will be a the last part of the review.

show abstract

Affinity chromatography of plasma proteins (guanidinobenzoatase): use of mimetic matrices and mimetic soluble ligands to prevent the binding of albumin on target affinity matrices

Cited by 6 publications

References 17 publications

Interaction of the Antitumor Drug 9-Aminoacridine with Guanidinobenzoatase Studied by Spectroscopic Methods: A Possible Tumor Marker Probe Based on the Fluorescence Exciplex Emission

Interaction of the Antitumor Drug 9-Aminoacridine with Guanidinobenzoatase Studied by Spectroscopic Methods: A Possible Tumor Marker Probe Based on the Fluorescence Exciplex Emission

Surface‐enhanced Raman and steady fluorescence study of interaction between antitumoral drug 9‐aminoacridine and trypsin‐like protease related to metastasis processes, guanidinobenzoatase

Strategies for the one-step immobilization–purification of enzymes as industrial biocatalysts

Contact Info

Product

Resources

About