Permeability of renal capillaries. I. Preparation of neutral and charged protein probes
This paper describes the preparation of charged and uncharged protein molecular probes for study of the permselectivity of renal capillaries. Horse heart myoglobin was used as a neutral myoglobin. Since it contained several fractions with different isoelectric points, it was purified by fast protein...
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crwiley:10.1111/j.1748-1716.1987.tb08070.x 2023-12-03T10:30:48+01:00 Permeability of renal capillaries. I. Preparation of neutral and charged protein probes ÖJTEG, G. NYGREN, K. WOLGAST, M. 1987 http://dx.doi.org/10.1111/j.1748-1716.1987.tb08070.x https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1748-1716.1987.tb08070.x https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1748-1716.1987.tb08070.x en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor Acta Physiologica Scandinavica volume 129, issue 3, page 277-286 ISSN 0001-6772 1365-201X Physiology journal-article 1987 crwiley https://doi.org/10.1111/j.1748-1716.1987.tb08070.x 2023-11-09T14:27:29Z This paper describes the preparation of charged and uncharged protein molecular probes for study of the permselectivity of renal capillaries. Horse heart myoglobin was used as a neutral myoglobin. Since it contained several fractions with different isoelectric points, it was purified by fast protein liquid chromatography (FPLC). To obtain a negatively charged myoglobin, the original horse heart myoglobin was treated with cyanate, resulting in net charge of ‐5.7 ± 0.3 at physiological pH (mean ± SEM). The charge was determined from the Donnan potential which develops over a semipermeable membrane separating the inside solution in which the protein was dissolved from a surrounding bath of equal ionic strength. Sperm whale myoglobin was similarly purified by FPLC and used as a positively (+ 1.7 ± 0.2) charged isomer. Horseradish peroxidase (HRP) was purified by means of gel and ion‐exchange chromatography and found to be neutral at physiological pH. Negatively charged (‐14.0 ± 0.5) HRP was obtained by succinylation. Two isomers of lactate dehyrogenase (LDH) were used, namely the slightly positive (+ 2) LDH‐M 4 and the strongly negative (‐19) LDH‐H 4 . These isomers, which occur naturally, did not require further purification. The Stokes‐Einstein radii, as measured by gel chromatography, of inulin, myoglobin, HRP and LDH were 11,17±5, 32 and 46 Å, respectively. The chemical modifications did not alter the Stokes‐Einstein radii. In biological studies on rat kidneys samples of both plasma and renal hilar lymph were found to contain radioactive low molecular weight degradation products in addition to the intact proteins. This necessitated separation of all individual samples on small Sephadex columns prior to analysis. Article in Journal/Newspaper Sperm whale Wiley Online Library (via Crossref) Acta Physiologica Scandinavica 129 3 277 286 |
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Wiley Online Library (via Crossref) |
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English |
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Physiology |
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Physiology ÖJTEG, G. NYGREN, K. WOLGAST, M. Permeability of renal capillaries. I. Preparation of neutral and charged protein probes |
topic_facet |
Physiology |
description |
This paper describes the preparation of charged and uncharged protein molecular probes for study of the permselectivity of renal capillaries. Horse heart myoglobin was used as a neutral myoglobin. Since it contained several fractions with different isoelectric points, it was purified by fast protein liquid chromatography (FPLC). To obtain a negatively charged myoglobin, the original horse heart myoglobin was treated with cyanate, resulting in net charge of ‐5.7 ± 0.3 at physiological pH (mean ± SEM). The charge was determined from the Donnan potential which develops over a semipermeable membrane separating the inside solution in which the protein was dissolved from a surrounding bath of equal ionic strength. Sperm whale myoglobin was similarly purified by FPLC and used as a positively (+ 1.7 ± 0.2) charged isomer. Horseradish peroxidase (HRP) was purified by means of gel and ion‐exchange chromatography and found to be neutral at physiological pH. Negatively charged (‐14.0 ± 0.5) HRP was obtained by succinylation. Two isomers of lactate dehyrogenase (LDH) were used, namely the slightly positive (+ 2) LDH‐M 4 and the strongly negative (‐19) LDH‐H 4 . These isomers, which occur naturally, did not require further purification. The Stokes‐Einstein radii, as measured by gel chromatography, of inulin, myoglobin, HRP and LDH were 11,17±5, 32 and 46 Å, respectively. The chemical modifications did not alter the Stokes‐Einstein radii. In biological studies on rat kidneys samples of both plasma and renal hilar lymph were found to contain radioactive low molecular weight degradation products in addition to the intact proteins. This necessitated separation of all individual samples on small Sephadex columns prior to analysis. |
format |
Article in Journal/Newspaper |
author |
ÖJTEG, G. NYGREN, K. WOLGAST, M. |
author_facet |
ÖJTEG, G. NYGREN, K. WOLGAST, M. |
author_sort |
ÖJTEG, G. |
title |
Permeability of renal capillaries. I. Preparation of neutral and charged protein probes |
title_short |
Permeability of renal capillaries. I. Preparation of neutral and charged protein probes |
title_full |
Permeability of renal capillaries. I. Preparation of neutral and charged protein probes |
title_fullStr |
Permeability of renal capillaries. I. Preparation of neutral and charged protein probes |
title_full_unstemmed |
Permeability of renal capillaries. I. Preparation of neutral and charged protein probes |
title_sort |
permeability of renal capillaries. i. preparation of neutral and charged protein probes |
publisher |
Wiley |
publishDate |
1987 |
url |
http://dx.doi.org/10.1111/j.1748-1716.1987.tb08070.x https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1748-1716.1987.tb08070.x https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1748-1716.1987.tb08070.x |
genre |
Sperm whale |
genre_facet |
Sperm whale |
op_source |
Acta Physiologica Scandinavica volume 129, issue 3, page 277-286 ISSN 0001-6772 1365-201X |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1111/j.1748-1716.1987.tb08070.x |
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Acta Physiologica Scandinavica |
container_volume |
129 |
container_issue |
3 |
container_start_page |
277 |
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286 |
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1784256841748840448 |