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When hyaluronan synthase from Pasteurella multocida was expressed in E. coli, the bacteria spun out a capsule of hyaluronan. This reaction was analyzed using purified membrane fragments containing the hyaluronan synthase, a radio-labeled tetrasaccharide acceptor (Part A), along with the activated forms of the sugar subunits: UDP-N-acetylglucosamine and UDP-glucuronic acid. In short incubations with the complete mixture, the tetrasaccharide primer elongated by several units, although only odd-numbered chains were visible (Part B lane 2). If one or both activated sugars were left out, the results shown in lanes 3-5 (Part B) were obtained. Why do you suppose that only odd-number chains are visible in your assay? (A) ACCEPTOR nonreducing end HO HO- OH COO™ O glucuronic acid (GICUA) HO- CH3 C=O NH CH₂ T OH O N-acetylglucosamine (GlcNAc) COO но- HO OH OH CH₂ reducing end NH T C=O CH₂ Choose one: O A. Synthase adds GlcNAc much more slowly than it adds GICUA. O B. One of the monosaccharide subunits (GlcNAc) is not labeled. OH O C. Addition of GICUA removes the radio label from the acceptor. O D. The enzyme adds only disaccharide units to the growing chain. (B) BIOSYNTHESIS number of residues 57913 origin |||| markers + GlcNAc + GICUA + GlcNAc 1 2 3 4 lane number (Part B from P.L. DeAngelis, J. Biol. Chem. 274:26557-26562, 1999. With permission from American Society for Biochemistry and Molecular Biology.) + GICUA 5 none

When hyaluronan synthase from Pasteurella multocida was expressed in E. coli, the bacteria spun out a capsule of hyaluronan. This reaction was analyzed using purified membrane fragments containing the hyaluronan synthase, a radio-labeled tetrasaccharide acceptor (Part A), along with the activated forms of the sugar subunits: UDP-N-acetylglucosamine and UDP-glucuronic acid. In short incubations with the complete mixture, the tetrasaccharide primer elongated by several units, although only odd-numbered chains were visible (Part B lane 2). If one or both activated sugars were left out, the results shown in lanes 3-5 (Part B) were obtained. Why do you suppose that only odd-number chains are visible in your assay? (A) ACCEPTOR nonreducing end HO HO- OH COO™ O glucuronic acid (GICUA) HO- CH3 C=O NH CH₂ T OH O N-acetylglucosamine (GlcNAc) COO но- HO OH OH CH₂ reducing end NH T C=O CH₂ Choose one: O A. Synthase adds GlcNAc much more slowly than it adds GICUA. O B. One of the monosaccharide subunits (GlcNAc) is not labeled. OH O C. Addition of GICUA removes the radio label from the acceptor. O D. The enzyme adds only disaccharide units to the growing chain. (B) BIOSYNTHESIS number of residues 57913 origin |||| markers + GlcNAc + GICUA + GlcNAc 1 2 3 4 lane number (Part B from P.L. DeAngelis, J. Biol. Chem. 274:26557-26562, 1999. With permission from American Society for Biochemistry and Molecular Biology.) + GICUA 5 none

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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When hyaluronan synthase from Pasteurella multocida was expressed in E. coli, the bacteria spun out a capsule of hyaluronan. This reaction was analyzed using purified membrane fragments containing the hyaluronan synthase, a radio-labeled tetrasaccharide acceptor (Part A), along with the activated forms of the sugar subunits: UDP-N-acetylglucosamine and UDP-glucuronic acid. In short incubations with the complete mixture, the tetrasaccharide primer elongated by several units, although only odd-numbered chains were visible (Part B lane 2). If one or both activated sugars were left out, the results shown in lanes 3-5 (Part B) were obtained. Why do you suppose that only odd-number chains are visible in your assay? (A) ACCEPTOR nonreducing end HO- HO OH COO glucuronic acid (GICUA) HO- CH3 T C=O I NH CH₂ OH N-acetylglucosamine (GlcNAc) COO но- HO OH OH I CH₂ reducing end NH C=O CH3 Choose one: O A. Synthase adds GlcNAc much more slowly than it adds GICUA. O B. One of the monosaccharide subunits (GlcNAc) is not labeled. OH O C. Addition of GICUA removes the radio label from the acceptor. O D. The enzyme adds only disaccharide units to the growing chain. (B) BIOSYNTHESIS number of residues 51953 origin ||||| markers + GICUA + GlcNAc + GlcNAc 1 2 3 4 lane number (Part B from P.L. DeAngelis, J. Biol. Chem. 274:26557-26562, 1999. With permission from American Society for Biochemistry and Molecular Biology.) + GICUA none 5
Transcribed Image Text:When hyaluronan synthase from Pasteurella multocida was expressed in E. coli, the bacteria spun out a capsule of hyaluronan. This reaction was analyzed using purified membrane fragments containing the hyaluronan synthase, a radio-labeled tetrasaccharide acceptor (Part A), along with the activated forms of the sugar subunits: UDP-N-acetylglucosamine and UDP-glucuronic acid. In short incubations with the complete mixture, the tetrasaccharide primer elongated by several units, although only odd-numbered chains were visible (Part B lane 2). If one or both activated sugars were left out, the results shown in lanes 3-5 (Part B) were obtained. Why do you suppose that only odd-number chains are visible in your assay? (A) ACCEPTOR nonreducing end HO- HO OH COO glucuronic acid (GICUA) HO- CH3 T C=O I NH CH₂ OH N-acetylglucosamine (GlcNAc) COO но- HO OH OH I CH₂ reducing end NH C=O CH3 Choose one: O A. Synthase adds GlcNAc much more slowly than it adds GICUA. O B. One of the monosaccharide subunits (GlcNAc) is not labeled. OH O C. Addition of GICUA removes the radio label from the acceptor. O D. The enzyme adds only disaccharide units to the growing chain. (B) BIOSYNTHESIS number of residues 51953 origin ||||| markers + GICUA + GlcNAc + GlcNAc 1 2 3 4 lane number (Part B from P.L. DeAngelis, J. Biol. Chem. 274:26557-26562, 1999. With permission from American Society for Biochemistry and Molecular Biology.) + GICUA none 5
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