Journal article
High-performance liquid chromatographic mapping and structural characterization of neurophysin isoforms
Analytical biochemistry, v 143(2), pp 215-225
1984
PMID: 6532238
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Both ion-exchange and reverse-phase HPLC protocols for micromapping of neurophysins have been examined and the structural relationships among the major isoforms identified in the maps have been characterized. Reverse-phase HPLC was found to be especially useful for obtaining fingerprints of the isoforms within each of the two major families of neurophysins, I (oxytocin-related) and II (vasopressin-related), for both bovine and human neurophysins from posterior pituitary sources. From fractionation of the bovine proteins on octylsilyl columns, at least four neurophysins I were identified, one of which corresponds to the intact sequence of 93 residues and three of which vary from the parent by various degrees of carboxyl-terminal truncation. For bovine neurophysin II, two isoforms were identified in the reverse-phase HPLC maps, both of 95 residues, which vary from one another by the residue, either Ile or Val, at position 89. Isoforms were also detected for human neurophysins, including a carboxyl-terminal truncated form of human neurophysin II. All of the major neurophysin isoforms and several of the minor forms were shown to be functionally active as expressed by their binding to peptide ligand affinity matrices. Reverse-phase HPLC mapping on the octylsilyl matrix allowed neurophysin fingerprinting of crude tissue extracts by providing a narrow “window” within which the neurophysins elute but many other polypeptides expected to be present are excluded. The reverse phase HPLC method provides a useful way to obtain isolated neurophysin isoforms for physicochemical characterizations now usually carried out with mixtures of isoforms obtained by ion-exchange chromatography. The method also has characteristics amenable both for high-sensitivity fingerprinting of neurophysin isoforms, from different species and anatomical sources, and as a prelude to microstructural and -functional characterization of the isoforms so isolated.
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Details
- Title
- High-performance liquid chromatographic mapping and structural characterization of neurophysin isoforms
- Creators
- Irwin M. Chaiken - National Institute of Diabetes and Digestive and Kidney DiseasesTed H. Miller - National Institute of Diabetes and Digestive and Kidney DiseasesReginald P. Sequeira - National Institute of Diabetes and Digestive and Kidney DiseasesTatsuhiko Kanmera - National Institute of Diabetes and Digestive and Kidney Diseases
- Publication Details
- Analytical biochemistry, v 143(2), pp 215-225
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology; Drexel University
- Web of Science ID
- WOS:A1984TZ96900001
- Scopus ID
- 2-s2.0-0021672759
- Other Identifier
- 991019520422804721
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InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology
- Chemistry, Analytical