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Kinetics of conformational changes in tRNA Phe (yeast) as studied by the fluorescence of the y-base and of formycin substituted for the 3'-terminal adenine
The kinetics of the melting transitions of tRNA phe (yeast) were followed by the fluorescence of the Y-base and of formycin substituted for the 3'-terminal adenine. As judged from differential UV absorbance melting cutves the formycin label had virtually no influence on the conformation of the...
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| Published in: | Biophysical chemistry 1975-10, Vol.3 (4), p.275-289 |
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| Language: | English |
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| container_end_page | 289 |
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| container_title | Biophysical chemistry |
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| creator | Coutts, Stephen M. Riesner, Detlev Römer, Roland Rabl, Cad R. Maass, Guenter |
| description | The kinetics of the melting transitions of tRNA
phe (yeast) were followed by the fluorescence of the Y-base and of formycin substituted for the 3'-terminal adenine. As judged from differential UV absorbance melting cutves the formycin label had virtually no influence on the conformation of the tRNA. A temperature jump apparatus was modified to allow the simultaneous observation of transmission and fluorescence intensities by two independent optical channels. The design of a temperature jump cell with an all quartz center piece is given. The cell is resistant to temperatures up to 90°C; it provides high optical sensitivity, low stray light intensity and the possibility of measuring fluorescence polarization. The
T-jump experiments allowed to discriminate between fast unspecific fluorescence quenching (τ |
| doi_str_mv | 10.1016/0301-4622(75)80020-2 |
| format | article |
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phe (yeast) were followed by the fluorescence of the Y-base and of formycin substituted for the 3'-terminal adenine. As judged from differential UV absorbance melting cutves the formycin label had virtually no influence on the conformation of the tRNA. A temperature jump apparatus was modified to allow the simultaneous observation of transmission and fluorescence intensities by two independent optical channels. The design of a temperature jump cell with an all quartz center piece is given. The cell is resistant to temperatures up to 90°C; it provides high optical sensitivity, low stray light intensity and the possibility of measuring fluorescence polarization. The
T-jump experiments allowed to discriminate between fast unspecific fluorescence quenching (τ <5 μsec) and slow co-operative conformational changes. In the central part of the temperature range of UV-melung (midpoint temperature 30°C in 0.01 M Na
+ and 39°C in 0.03 M Na
+, pH 6.8) two resolvable relaxation processes were observed. The coirssponding relaxation times were 20 msec and 800 msec at 30°C in 0.01 M Na
+, and 4 msec and 120 msec at 39°C in 0.03 M Na
+. The Y-base fluorescence shows both of the relaxation effects, which almost cancel in equilibrium fluorescence melting, because their amplitudes have opposite signs. From this finding the existence of some residual tertiary structure is inferred which persists after the unfolding of the main part of tertiary structure durirg early melting (midpoint temperature 24°C in 0.03 M Na
+). In the fluorescence sigXXX of the formycin also the two relaxation effects appear. Both of them are connected with a decrease of the fluorescence intensity. From the results a coupled opening of the anticodon and acceptor branches is concluded.
Enzymes: phenylalanyl-tRNA synthetase, PRS (EC 6.1.1.-20); ATP (CTP) tRNA nucleotidyl transferase, NT (EC 2.7.7.-20); alkaline phosphatase (EC 3-1-3.1).</description><identifier>ISSN: 0301-4622</identifier><identifier>EISSN: 1873-4200</identifier><identifier>DOI: 10.1016/0301-4622(75)80020-2</identifier><identifier>PMID: 1103985</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Adenine ; Binding Sites ; Formycins ; Hot Temperature ; Kinetics ; Mathematics ; Nucleic Acid Conformation ; Nucleic Acid Denaturation ; Phenylalanine ; RNA Nucleotidyltransferases ; RNA, Transfer ; Saccharomyces cerevisiae ; Spectrometry, Fluorescence ; Spectrophotometry, Ultraviolet</subject><ispartof>Biophysical chemistry, 1975-10, Vol.3 (4), p.275-289</ispartof><rights>1975</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/0301462275800202$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3619,27924,27925,45983</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1103985$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Coutts, Stephen M.</creatorcontrib><creatorcontrib>Riesner, Detlev</creatorcontrib><creatorcontrib>Römer, Roland</creatorcontrib><creatorcontrib>Rabl, Cad R.</creatorcontrib><creatorcontrib>Maass, Guenter</creatorcontrib><title>Kinetics of conformational changes in tRNA Phe (yeast) as studied by the fluorescence of the y-base and of formycin substituted for the 3'-terminal adenine</title><title>Biophysical chemistry</title><addtitle>Biophys Chem</addtitle><description>The kinetics of the melting transitions of tRNA
phe (yeast) were followed by the fluorescence of the Y-base and of formycin substituted for the 3'-terminal adenine. As judged from differential UV absorbance melting cutves the formycin label had virtually no influence on the conformation of the tRNA. A temperature jump apparatus was modified to allow the simultaneous observation of transmission and fluorescence intensities by two independent optical channels. The design of a temperature jump cell with an all quartz center piece is given. The cell is resistant to temperatures up to 90°C; it provides high optical sensitivity, low stray light intensity and the possibility of measuring fluorescence polarization. The
T-jump experiments allowed to discriminate between fast unspecific fluorescence quenching (τ <5 μsec) and slow co-operative conformational changes. In the central part of the temperature range of UV-melung (midpoint temperature 30°C in 0.01 M Na
+ and 39°C in 0.03 M Na
+, pH 6.8) two resolvable relaxation processes were observed. The coirssponding relaxation times were 20 msec and 800 msec at 30°C in 0.01 M Na
+, and 4 msec and 120 msec at 39°C in 0.03 M Na
+. The Y-base fluorescence shows both of the relaxation effects, which almost cancel in equilibrium fluorescence melting, because their amplitudes have opposite signs. From this finding the existence of some residual tertiary structure is inferred which persists after the unfolding of the main part of tertiary structure durirg early melting (midpoint temperature 24°C in 0.03 M Na
+). In the fluorescence sigXXX of the formycin also the two relaxation effects appear. Both of them are connected with a decrease of the fluorescence intensity. From the results a coupled opening of the anticodon and acceptor branches is concluded.
Enzymes: phenylalanyl-tRNA synthetase, PRS (EC 6.1.1.-20); ATP (CTP) tRNA nucleotidyl transferase, NT (EC 2.7.7.-20); alkaline phosphatase (EC 3-1-3.1).</description><subject>Adenine</subject><subject>Binding Sites</subject><subject>Formycins</subject><subject>Hot Temperature</subject><subject>Kinetics</subject><subject>Mathematics</subject><subject>Nucleic Acid Conformation</subject><subject>Nucleic Acid Denaturation</subject><subject>Phenylalanine</subject><subject>RNA Nucleotidyltransferases</subject><subject>RNA, Transfer</subject><subject>Saccharomyces cerevisiae</subject><subject>Spectrometry, Fluorescence</subject><subject>Spectrophotometry, Ultraviolet</subject><issn>0301-4622</issn><issn>1873-4200</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1975</creationdate><recordtype>article</recordtype><recordid>eNo9UctO3TAUtFArernlD6jkVYFFWj-u42SDhBB9qAiqqqytE_ukuEociJ1K-Zb-bG24qjeWZ8ZzHkPICWcfOOP1RyYZr3a1EGdanTeMCVaJA7LhjZbVTjD2imz-S96Qoxh_s3yy8JAccs5k26gN-fvNB0zeRjr11E6hn-YRkp8CDNQ-QPiFkfpA04_bS_r9AenZihDTOYVIY1qcR0e7labM9MMyzRgtBovFrGBr1UFECsEVpHivNrvFpYvJpyXl3xl8lsrTKuE8-lIYHIbc1lvyuoch4vH-3pL7T9c_r75UN3efv15d3lTIWaMqC64VCgVaUAqc24Hs27YBW6ua6065PL9THNVOa6mYAKE05LfTsleS9XJL3r_4Ps7T04IxmdHnOYYBAk5LNI1oa63yWrfk3V64dCM68zj7EebV7LeZ-YsXHnO3fzzOJlpf9uH8jDYZN3nDmSnpmRKNKdEYrcxzekbIfxrujA8</recordid><startdate>197510</startdate><enddate>197510</enddate><creator>Coutts, Stephen M.</creator><creator>Riesner, Detlev</creator><creator>Römer, Roland</creator><creator>Rabl, Cad R.</creator><creator>Maass, Guenter</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>197510</creationdate><title>Kinetics of conformational changes in tRNA Phe (yeast) as studied by the fluorescence of the y-base and of formycin substituted for the 3'-terminal adenine</title><author>Coutts, Stephen M. ; Riesner, Detlev ; Römer, Roland ; Rabl, Cad R. ; Maass, Guenter</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-e1085-cad925e2eca55add4a3f998ac65617b5d622d51e54773502a257a51ed73f530f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1975</creationdate><topic>Adenine</topic><topic>Binding Sites</topic><topic>Formycins</topic><topic>Hot Temperature</topic><topic>Kinetics</topic><topic>Mathematics</topic><topic>Nucleic Acid Conformation</topic><topic>Nucleic Acid Denaturation</topic><topic>Phenylalanine</topic><topic>RNA Nucleotidyltransferases</topic><topic>RNA, Transfer</topic><topic>Saccharomyces cerevisiae</topic><topic>Spectrometry, Fluorescence</topic><topic>Spectrophotometry, Ultraviolet</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Coutts, Stephen M.</creatorcontrib><creatorcontrib>Riesner, Detlev</creatorcontrib><creatorcontrib>Römer, Roland</creatorcontrib><creatorcontrib>Rabl, Cad R.</creatorcontrib><creatorcontrib>Maass, Guenter</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Biophysical chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Coutts, Stephen M.</au><au>Riesner, Detlev</au><au>Römer, Roland</au><au>Rabl, Cad R.</au><au>Maass, Guenter</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Kinetics of conformational changes in tRNA Phe (yeast) as studied by the fluorescence of the y-base and of formycin substituted for the 3'-terminal adenine</atitle><jtitle>Biophysical chemistry</jtitle><addtitle>Biophys Chem</addtitle><date>1975-10</date><risdate>1975</risdate><volume>3</volume><issue>4</issue><spage>275</spage><epage>289</epage><pages>275-289</pages><issn>0301-4622</issn><eissn>1873-4200</eissn><abstract>The kinetics of the melting transitions of tRNA
phe (yeast) were followed by the fluorescence of the Y-base and of formycin substituted for the 3'-terminal adenine. As judged from differential UV absorbance melting cutves the formycin label had virtually no influence on the conformation of the tRNA. A temperature jump apparatus was modified to allow the simultaneous observation of transmission and fluorescence intensities by two independent optical channels. The design of a temperature jump cell with an all quartz center piece is given. The cell is resistant to temperatures up to 90°C; it provides high optical sensitivity, low stray light intensity and the possibility of measuring fluorescence polarization. The
T-jump experiments allowed to discriminate between fast unspecific fluorescence quenching (τ <5 μsec) and slow co-operative conformational changes. In the central part of the temperature range of UV-melung (midpoint temperature 30°C in 0.01 M Na
+ and 39°C in 0.03 M Na
+, pH 6.8) two resolvable relaxation processes were observed. The coirssponding relaxation times were 20 msec and 800 msec at 30°C in 0.01 M Na
+, and 4 msec and 120 msec at 39°C in 0.03 M Na
+. The Y-base fluorescence shows both of the relaxation effects, which almost cancel in equilibrium fluorescence melting, because their amplitudes have opposite signs. From this finding the existence of some residual tertiary structure is inferred which persists after the unfolding of the main part of tertiary structure durirg early melting (midpoint temperature 24°C in 0.03 M Na
+). In the fluorescence sigXXX of the formycin also the two relaxation effects appear. Both of them are connected with a decrease of the fluorescence intensity. From the results a coupled opening of the anticodon and acceptor branches is concluded.
Enzymes: phenylalanyl-tRNA synthetase, PRS (EC 6.1.1.-20); ATP (CTP) tRNA nucleotidyl transferase, NT (EC 2.7.7.-20); alkaline phosphatase (EC 3-1-3.1).</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>1103985</pmid><doi>10.1016/0301-4622(75)80020-2</doi><tpages>15</tpages></addata></record> |
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| identifier | ISSN: 0301-4622 |
| ispartof | Biophysical chemistry, 1975-10, Vol.3 (4), p.275-289 |
| issn | 0301-4622 1873-4200 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_82967587 |
| source | ScienceDirect Physical & Analytical Chemistry Backfile |
| subjects | Adenine Binding Sites Formycins Hot Temperature Kinetics Mathematics Nucleic Acid Conformation Nucleic Acid Denaturation Phenylalanine RNA Nucleotidyltransferases RNA, Transfer Saccharomyces cerevisiae Spectrometry, Fluorescence Spectrophotometry, Ultraviolet |
| title | Kinetics of conformational changes in tRNA Phe (yeast) as studied by the fluorescence of the y-base and of formycin substituted for the 3'-terminal adenine |
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