Isolation by zinc-affinity chromatography of the histidine–proline-rich-glycoprotein molecule associated with rabbit skeletal muscle AMP deaminase

التفاصيل البيبلوغرافية
العنوان: Isolation by zinc-affinity chromatography of the histidine–proline-rich-glycoprotein molecule associated with rabbit skeletal muscle AMP deaminase
المؤلفون: Antonietta Raffaella Maria Sabbatini, Antonio Raggi, Maria Ranieri-Raggi, Arthur J. G. Moir, Daniela Martini
المصدر: Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 1645:81-88
بيانات النشر: Elsevier BV, 2003.
سنة النشر: 2003
مصطلحات موضوعية: chemistry.chemical_classification, Protein subunit, Biophysics, AMP deaminase, Biochemistry, Analytical Chemistry, N-terminus, Enzyme, chemistry, Affinity chromatography, Binding site, Glycoprotein, Molecular Biology, Histidine
الوصف: The histidine-proline-rich glycoprotein (HPRG) component of rabbit skeletal muscle AMP deaminase under denaturing and reducing conditions specifically binds to a Zn(2+)-charged affinity column and is only eluted with an EDTA-containing buffer that strips Zn(2+) from the gel. The isolated protein is homogeneous showing an apparent molecular weight (MW) of 95000 and the N-terminal sequence L-T-P-T-D-X-K-T-T-K-P-L-A-E-K-A-L-D-L-I, corresponding to that of rabbit plasma HPRG. The incubation with peptide-N-glycosidase F promotes the reduction of the apparent MW of isolated HPRG to 70000, characterizing it as a N-glycosylated protein. The separation from AMP deaminase of an 85-kDa component with a blocked N terminus is observed when the enzyme is applied to the Zn-charged column under nondenaturing conditions. On storage under reducing conditions, this component undergoes an 85- to 95-kDa transition yielding a L-T-P-T-D-X-K-T-T-K-P-L N-terminal sequence, suggesting that the shift in the migration on SDS/PAGE as well as the truncation of the protein at its N terminus are promoted by the reduction of a disulfide bond present in freshly isolated HPRG. The separation of HPRG induces a marked reduction in the solubility of AMP deaminase, strongly suggesting a role of HPRG in assuring the molecular integrity of the enzyme.
تدمد: 1570-9639
DOI: 10.1016/s1570-9639(02)00527-7
URL الوصول: https://explore.openaire.eu/search/publication?articleId=doi_________::483808b2517b88fa91cd2df3b2b58220
https://doi.org/10.1016/s1570-9639(02)00527-7
Rights: CLOSED
رقم الانضمام: edsair.doi...........483808b2517b88fa91cd2df3b2b58220
قاعدة البيانات: OpenAIRE
الوصف
تدمد:15709639
DOI:10.1016/s1570-9639(02)00527-7