Structural characterization of insulin NPH formulations
(2007) In European Journal of Pharmaceutical Sciences 30(5). p.414-423- Abstract
- Insulin NPH (neutral protamine hagedorn) has for long been one of the most important therapeutic formulations for the treatment of diabetes. The protracted action profile of NPH formulations is gained from crystallizing insulin with zinc in the presence of the basic poly-arginine peptide protamine. In spite of its long history and successful use, the binding mode of the insulin-protamine complex is not known. in this study, three different systems were used to study protamine binding to insulin. In the first system, crystals of an insulin-protamine complex grown in the presence of urea and diffracting to 1.5 angstrom resolution were analyzed. In the second system, a shorter peptide consisting of 12 arginine residues was co-crystallized... (More)
- Insulin NPH (neutral protamine hagedorn) has for long been one of the most important therapeutic formulations for the treatment of diabetes. The protracted action profile of NPH formulations is gained from crystallizing insulin with zinc in the presence of the basic poly-arginine peptide protamine. In spite of its long history and successful use, the binding mode of the insulin-protamine complex is not known. in this study, three different systems were used to study protamine binding to insulin. In the first system, crystals of an insulin-protamine complex grown in the presence of urea and diffracting to 1.5 angstrom resolution were analyzed. In the second system, a shorter peptide consisting of 12 arginine residues was co-crystallized with insulin in order to reduce the flexibility and thereby improve the electron density of the peptide. Both systems yielded data to a significantly higher resolution than obtained previously. In addition, a third system was analyzed where crystals of insulin and protamine were grown in the absence of urea, with conditions closely resembling the pharmaceutical formulation. Data from these NPH microcrystals could for the first time be collected to 2.2 angstrom resolution at a micro focused X-ray beamline. Analysis of all three crystal forms reveal potential protamine density located close to the solvent channel leading to the centrally located zinc atoms in the insulin hexamer and support that protamine binds to insulin in a not well defined conformation. (c) 2007 Elsevier B.V. All rights reserved. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/664953
- author
- Norrman, Mathias LU ; Hubalek, Frantisek and Schluckebier, Gerd
- organization
- publishing date
- 2007
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- protamine, NPH (neutral protamine hagedorn), insulin, X-ray, microcrystal
- in
- European Journal of Pharmaceutical Sciences
- volume
- 30
- issue
- 5
- pages
- 414 - 423
- publisher
- Elsevier
- external identifiers
-
- wos:000245816000007
- scopus:33947261481
- pmid:17339105
- ISSN
- 1879-0720
- DOI
- 10.1016/j.ejps.2007.01.003
- language
- English
- LU publication?
- yes
- id
- 1e3c577c-f74a-4390-b4e6-1b99aa0db3aa (old id 664953)
- date added to LUP
- 2016-04-01 11:42:03
- date last changed
- 2022-01-26 08:56:05
@article{1e3c577c-f74a-4390-b4e6-1b99aa0db3aa,
abstract = {{Insulin NPH (neutral protamine hagedorn) has for long been one of the most important therapeutic formulations for the treatment of diabetes. The protracted action profile of NPH formulations is gained from crystallizing insulin with zinc in the presence of the basic poly-arginine peptide protamine. In spite of its long history and successful use, the binding mode of the insulin-protamine complex is not known. in this study, three different systems were used to study protamine binding to insulin. In the first system, crystals of an insulin-protamine complex grown in the presence of urea and diffracting to 1.5 angstrom resolution were analyzed. In the second system, a shorter peptide consisting of 12 arginine residues was co-crystallized with insulin in order to reduce the flexibility and thereby improve the electron density of the peptide. Both systems yielded data to a significantly higher resolution than obtained previously. In addition, a third system was analyzed where crystals of insulin and protamine were grown in the absence of urea, with conditions closely resembling the pharmaceutical formulation. Data from these NPH microcrystals could for the first time be collected to 2.2 angstrom resolution at a micro focused X-ray beamline. Analysis of all three crystal forms reveal potential protamine density located close to the solvent channel leading to the centrally located zinc atoms in the insulin hexamer and support that protamine binds to insulin in a not well defined conformation. (c) 2007 Elsevier B.V. All rights reserved.}},
author = {{Norrman, Mathias and Hubalek, Frantisek and Schluckebier, Gerd}},
issn = {{1879-0720}},
keywords = {{protamine; NPH (neutral protamine hagedorn); insulin; X-ray; microcrystal}},
language = {{eng}},
number = {{5}},
pages = {{414--423}},
publisher = {{Elsevier}},
series = {{European Journal of Pharmaceutical Sciences}},
title = {{Structural characterization of insulin NPH formulations}},
url = {{http://dx.doi.org/10.1016/j.ejps.2007.01.003}},
doi = {{10.1016/j.ejps.2007.01.003}},
volume = {{30}},
year = {{2007}},
}