Isolation and characterization of ovine IGFBP-4: protein purification and cDNA sequence

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dc.contributor.authorCARR, J.M.
dc.contributor.authorGRANT, P.A.
dc.contributor.authorFRANCIS, G.L.
dc.contributor.authorOWENS, J.A.
dc.contributor.authorWALLACE, J.C.
dc.contributor.authorWALTON, P.E.
dc.date.issued1994
dc.description.abstractThree different molecular mass forms of IGF-binding proteins (IGFBPs) were purified from ovine plasma by IGF-I affinity chromatography and reverse-phase HPLC: a 46 kDa doublet and 29 kDa and 24 kDa forms. Amino-terminal sequence analysis confirmed that these proteins were ovine (o)IGFBP-3 (46 kDa) and two molecular size variants of oIGFBP-4. oIGFBP-3 and the 29 kDa form of oIGFBP-4 were shown to be N-glycosylated. Isoelectric points were determined to be at approximately pH 6 for oIGFBP-3 and at pH 7 and pH 7.5 for the 29 and 24 kDa forms of oIGFBP-4 respectively. The two different molecular mass variants of oIGFBP-4 had similar IGF-binding properties. Compared with human IGFBP-3 and oIGFBP-3, the two variants of oIGFBP-4 exhibited lower relative binding to amino-terminally modified IGF-I analogues in a competitive IGF-binding assay. The full protein sequence of oIGFBP-4, as deduced from the cDNA sequence, showed a high degree of identity with rat (90%), human (96%) and bovine (98%) IGFBP-4. The cDNA sequence also showed homology over regions of the 3' non-coding sequence, particularly in comparison with bovine IGFBP-4 (96%). Northern analysis of mRNA for oIGFBP-4 indicated a 2.6 kb major transcript and two minor transcripts of approximately 2.1 and 1.8 kb. oIGFBP-4 mRNA transcripts were detected in adult ewe liver > kidney > lung >> heart and also in several fetal tissues, thus suggesting tissue-specific and developmental regulation. The availability of purified oIGFBP-4 and oIGFBP-3 as well as DNA probes for oIGFBP-4 will enable further study of the properties and functions of these proteins, as well as the establishment of specific assays for these IGFBPs.
dc.description.statementofresponsibilityJ M Carr, P A Grant, G L Francis, J A Owens, J C Wallace and P E Walton
dc.identifier.citationJournal of Molecular Endocrinology, 1994; 13(2):219-236
dc.identifier.doi10.1677/jme.0.0130219
dc.identifier.issn1479-6813
dc.identifier.issn1479-6813
dc.identifier.orcidOWENS, J.A. [0000-0002-7498-1353]
dc.identifier.urihttp://hdl.handle.net/2440/92947
dc.language.isoen
dc.publisherBioScientifica
dc.rights©1994 Journal of Endocrinology
dc.source.urihttps://doi.org/10.1677/jme.0.0130219
dc.subjectAnimals
dc.subjectCattle
dc.subjectSheep
dc.subjectHumans
dc.subjectRats
dc.subjectSomatomedins
dc.subjectInsulin-Like Growth Factor Binding Proteins
dc.subjectInsulin-Like Growth Factor Binding Protein 4
dc.subjectCloning, Molecular
dc.subjectBase Sequence
dc.subjectSequence Homology, Amino Acid
dc.subjectGlycosylation
dc.subjectSpecies Specificity
dc.subjectAmino Acid Sequence
dc.subjectMolecular Sequence Data
dc.subjectDNA, Complementary
dc.subjectCarrier Proteins
dc.subjectMolecular Weight
dc.subjectRNA, Messenger
dc.titleIsolation and characterization of ovine IGFBP-4: protein purification and cDNA sequence
dc.typeJournal article
pubs.publication-statusPublished

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