The steroid progesterone has an important role in the femalereproductive system. Researchers interested in studying membraneprogestin receptors (MPRS) developed a method to produce and purifythe protein in active form.First, the researchers devised a way to prepare a specific MPRknown as hMPRA using the machinery of yeast cells. In order tofacilitate purification and identification in later studies, theymanipulated the yeast cells so that they attached two different tags tothe C-terminal end of the protein.The first tag, Compound 1, is a peptide sequence that acts as anepitope, part of a much larger peptide sequence that is recognized bythe immune system.The second sequence consisted of six consecutive histidineresidues (His). This sequence binds tightly to Ni2+ cations. Inchromatography, (His), tag labeled proteins can be eluted from Ni²+.supported columns by adding a small molecule to the eluent thatmimics the side chain of histidine.Glu-Gin-Lys-Leu-lle-Ser-Glu-Glu-Asp-LeuCompound 1After preparing hMPRA, the researchers conducted a binding assaythat utilized tritium labeled 1,2,6,7-[H³]-progesterone to measure the Kaof the receptor while still in the membranes of the yeast cells.hMPRA was then extracted from the membranes usingn-decyl-B-D-maltopyranoside, Compound 2. The researchers purified thehMPRA with two successive rounds of chromatography that exploitedeach of the tags. The buffers used to elute the protein contained300 mM NaCl, 50 mM NaH₂PO, (pK, = 7.2), and various amounts ofNaOH (MM 40 g/mol). During the first chromatography step, aspecific chemical agent was immobilized on the stationary phase tobind to the Compound 1 tag. After the second chromatography step,which utilized the (His), tag, the researchers used the same bindingassay and found that K, was similar.Adapted from M. B. Hossain, T. Oshima, S. Hirose, J. Wang, and T. Tokumoto, "Expression and Purificationof Human Membrane Progestin Receptor a (mPRA)." PLOS One. ©2015 Creative Commons Attribution.The second purification step is which type of chromatographicseparation?A. AffinityOB. Size exclusionO c. Cation exchangeO D. Anion exchangeThe structure of Compound 2 is shown.но.HOHOOHOОН"OHO A. 7.5 x 10-3OB. 3.0 x 10-3O c. 3.0 x 10²O D. 1.5 x10-¹OHCompound 2What structural feature(s) is(are) most important to the functioning ofthis compound as described in the passage?O A. Specific configuration of numerous chirality centersOB. Multiple hydrolysable linkagesC. Combination of large hydrophobic and hydrophilic regionsO D. Presence of a reducing sugarHow many moles of NaCl were contained in 500 mL of the buffersolution used to elute hMPRa?Which experimental evidence suggests that the purified hMPRaobtained by the researchers was in its native state?The hMPRA that was obtained:O A. retained both the Compound 1 and (His), tags.OB. was purified by two separate chromatography steps.O c. exhibited a binding affinity for progesterone that was similarto that exhibited by native hMPRA.OD. had a nearly identical molecular weight to hMPRA obtainedelsewhere.The ligand of hMPRA is derived from which compound?O A. GlucoseO B. PhenylalanineO c. GlycerolD. Cholesterol

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The steroid progesterone has an important role in the femalereproductive system. Researchers interested in studying membraneprogestin receptors (MPRS) developed a method to produce and purifythe protein in active form.First, the researchers devised a way to prepare a specific MPRknown as hMPRA using the machinery of yeast cells. In order tofacilitate purification and identification in later studies, theymanipulated the yeast cells so that they attached two different tags tothe C-terminal end of the protein.The first tag, Compound 1, is a peptide sequence that acts as anepitope, part of a much larger peptide sequence that is recognized bythe immune system.The second sequence consisted of six consecutive histidineresidues (His). This sequence binds tightly to Ni2+ cations. Inchromatography, (His), tag labeled proteins can be eluted from Ni²+.supported columns by adding a small molecule to the eluent thatmimics the side chain of histidine.Glu-Gin-Lys-Leu-lle-Ser-Glu-Glu-Asp-LeuCompound 1After preparing hMPRA, the researchers conducted a binding assaythat utilized tritium labeled 1,2,6,7-[H³]-progesterone to measure the Kaof the receptor while still in the membranes of the yeast cells.hMPRA was then extracted from the membranes usingn-decyl-B-D-maltopyranoside, Compound 2. The researchers purified thehMPRA with two successive rounds of chromatography that exploitedeach of the tags. The buffers used to elute the protein contained300 mM NaCl, 50 mM NaH₂PO, (pK, = 7.2), and various amounts ofNaOH (MM 40 g/mol). During the first chromatography step, aspecific chemical agent was immobilized on the stationary phase tobind to the Compound 1 tag. After the second chromatography step,which utilized the (His), tag, the researchers used the same bindingassay and found that K, was similar.Adapted from M. B. Hossain, T. Oshima, S. Hirose, J. Wang, and T. Tokumoto, &#34;Expression and Purificationof Human Membrane Progestin Receptor a (mPRA).&#34; PLOS One. ©2015 Creative Commons Attribution.The second purification step is which type of chromatographicseparation?A. AffinityOB. Size exclusionO c. Cation exchangeO D. Anion exchangeThe structure of Compound 2 is shown.но.HOHOOHOОН&#34;OHO A. 7.5 x 10-3OB. 3.0 x 10-3O c. 3.0 x 10²O D. 1.5 x10-¹OHCompound 2What structural feature(s) is(are) most important to the functioning ofthis compound as described in the passage?O A. Specific configuration of numerous chirality centersOB. Multiple hydrolysable linkagesC. Combination of large hydrophobic and hydrophilic regionsO D. Presence of a reducing sugarHow many moles of NaCl were contained in 500 mL of the buffersolution used to elute hMPRa?Which experimental evidence suggests that the purified hMPRaobtained by the researchers was in its native state?The hMPRA that was obtained:O A. retained both the Compound 1 and (His), tags.OB. was purified by two separate chromatography steps.O c. exhibited a binding affinity for progesterone that was similarto that exhibited by native hMPRA.OD. had a nearly identical molecular weight to hMPRA obtainedelsewhere.The ligand of hMPRA is derived from which compound?O A. GlucoseO B. PhenylalanineO c. GlycerolD. Cholesterol

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