@article{e7e417662d124cac9ce57c530ae56f54,
title = "Cloning and characterization of a bovine alpha interferon receptor",
abstract = "A bovine interferon α receptor (BoIFN-αR1) cDNA, homologous to the human cDNA [1], was isolated. Transfection of the BoIFN-αR1 cDNA into monkey COS cells results in a large increase in high-affinity binding sites for human IFN-αA and IFN-αB. Covalent crosslinking of radiolabeled HuIFN-αA and -αB demonstrates that the complex of [32P]HuIFN with the BoIFN-αR1 protein (predicted mass, 61375) expressed in COS cells migrates as a 140-150 kDa band.",
keywords = "Interferon receptor, Protein crosslinking, cDNA sequence, α-Interferon",
author = "Lim, {Jin Kyu} and Langer, {Jerome A.}",
note = "Funding Information: We have thus isolated and characterizead bovine cDNA encodinga functionaIl FN-a receptorh, omolo-gous to the previouslyc lonedhuman and murine IFN-aRls \[1,8\]T. owardthe end of our studies,a n almost identical bovine MDBK IFN-aR1 cDNA was de- scribed,a nd its ability,w hene xpresseodn humanc ells, to bind HuIFN-aA, -aB and -aD, was documented \[9,10\].T he protein sequences are in agreement: Mouchel-Vielhe t al. \[10\f]o und phenylalaninaet position 422 an their MDBK clone, as we found in the MDBK partial clones. When the BoIFN-aR1 cDNA was expressedi n monkeyC OS cells, the transfectecde llss howeda great enhancemenotf high-affinityb inding of HuIFN-aA (Fig. 2) and -aB (not shown).T he easeo f detectingth e transfecteBdo IFN-aR1 proteinb y ligand bindingc on-trasts with the difficulty in similarly detectingt he HuIFN-aR1, despitet he presenceo f large amountso f HuIFN-aR1 mRNA in these transfectecde lls. This is further evidencet hat the bovine IFN-aR1 may have some advantageosv er the human protein for the in vitro studyo f ligand-receptoinrt eractions. The large number of high-affinityb indingsites on COS cells transfectewd ith the BoIFN-aR1/pcDNAI plasmids uggeststh at the BolFN-aR1 is sufficientf or the binding of IFN-aA. If a putatives econdsubunit were required for binding, then this subunit would have to be present in 0.25-1.0.106c opies in COS cells, which seemsu nlikely.N everthelesss,in cevarious experimentws ith the human and murine IFN-aRls suggestth ata ccessorfya ctorsa re requiredo r moderate the bindingo f IFN-a subtype\s[ 1,8,11-13,36it\ ]w, ill be interesting to determine definitive\[yw hether the BoIFN-aR1 proteini s both necessarayn d sufficientf or IFN-a binding,o r whetherit requireso therfactors. Using SDS-PAGE, including 6.7 M urea \[31\]w, e resolveda novel 140-150k Da IFN/receptorc omplex in COS cells transfectewd ith the BoIFN-aR1. With a standardS DS-PAGE systeml ackingurea, we saw an enhancemenotf a broad band in the regiono f the band seen on untransfectecde lls,but were unablet o resolve the new band from the endogenouCs OS band; from gels lacking urea, we estimated the size of the crosslinkedc omplexa t 130-150k Da (data not shown), close to that determinedfr om the SDS-urea gels. We thus estimatet hat the expressedB oIFN-aR1 protein has an M r of 110-130000T. his is similar but not identicalt o the major proteinc ovalentlyc rosslinkedto radiolabeledIF N-aA or -aB on bovine cells (Fig. 4, 'MDBK'; Refs. 37-39). These experimentasr e the first determinatioonf the size of an expressedIF N-aR1 protein. We gratefullya cknowledgesu pportb y grants from the American Cancer Society (#BC-698) and the Foundationo f the UMDNJ (#14-92)t o J.A.L., and departmentafel llowships upportt o J.-K.L. We thank Dr. Gilles Uz6 for generouslys upplyinga clone con-tainingt he HuIFN-aR1 cDNA, from which the probes used initially for screeningw ere made. We thank our departmentaclo lleaguese, speciallyD rs. Indira Krish-nan, Robert Donnellya nd SidneyP estka.W e sincerely",
year = "1993",
month = jun,
day = "25",
doi = "10.1016/0167-4781(93)90129-2",
language = "English (US)",
volume = "1173",
pages = "314--319",
journal = "BBA - Gene Structure and Expression",
issn = "0167-4781",
publisher = "Elsevier BV",
number = "3",
}