Effects of Adenylyl Imidodiphosphate, a Nonhydrolyzable Adenosine Triphosphate Analog, on Reactivated and Rigor Wave Sea Urchin Sperm
A nonhydrolyzable ATP analog, adenylyl imidodiphosphate (AMP-PNP), has been used to study the role of ATP binding in flagellar motility. Sea urchin sperm of Lytechinus pictus were demembranated, reactivated, and locked in "rigor waves" by a modification of the method of Gibbons and Gibbons...
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| Veröffentlicht in: | The Journal of cell biology 1978-12, Vol.79 (3), p.827-832 |
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| container_title | The Journal of cell biology |
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| creator | Penningroth, Stephen M. Witman, George B. |
| description | A nonhydrolyzable ATP analog, adenylyl imidodiphosphate (AMP-PNP), has been used to study the role of ATP binding in flagellar motility. Sea urchin sperm of Lytechinus pictus were demembranated, reactivated, and locked in "rigor waves" by a modification of the method of Gibbons and Gibbons (11). Rigor wave sperm relaxed within 2 min after addition of 4 μM ATP, and reactivated upon addition of 10-12 μM ATP. The beat frequency of the reactivated sperm varied with ATP concentration according to Michaelis-Menten kinetics (`` Km''=0.24 mM;`` Vmax''=44 Hz) and was competitively inhibited by AMP-PNP (`` Ki''≅ 8.1 mM). Rigor wave sperm were completely relaxed (straightened) within 2 min by AMP-PNP at concentrations of 2-4 mM. The possibilities that relaxation in AMP-PNP was a result of ATP contamination, AMP-PNP hydrolysis, or lowering of the free Mg++concentration were conclusively ruled out. The results suggest that dynein cross-bridge release is dependent upon ATP binding but not hydrolysis. |
| doi_str_mv | 10.1083/jcb.79.3.827 |
| format | Article |
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Sea urchin sperm of Lytechinus pictus were demembranated, reactivated, and locked in "rigor waves" by a modification of the method of Gibbons and Gibbons (11). Rigor wave sperm relaxed within 2 min after addition of 4 μM ATP, and reactivated upon addition of 10-12 μM ATP. The beat frequency of the reactivated sperm varied with ATP concentration according to Michaelis-Menten kinetics (`` Km''=0.24 mM;`` Vmax''=44 Hz) and was competitively inhibited by AMP-PNP (`` Ki''≅ 8.1 mM). Rigor wave sperm were completely relaxed (straightened) within 2 min by AMP-PNP at concentrations of 2-4 mM. The possibilities that relaxation in AMP-PNP was a result of ATP contamination, AMP-PNP hydrolysis, or lowering of the free Mg++concentration were conclusively ruled out. The results suggest that dynein cross-bridge release is dependent upon ATP binding but not hydrolysis.</description><identifier>ISSN: 0021-9525</identifier><identifier>EISSN: 1540-8140</identifier><identifier>DOI: 10.1083/jcb.79.3.827</identifier><identifier>PMID: 153347</identifier><language>eng</language><publisher>United States: Rockefeller University Press</publisher><subject>Adenosine triphosphatases ; Adenosine Triphosphate - analogs & derivatives ; Adenosine Triphosphate - metabolism ; Adenosine Triphosphate - pharmacology ; Adenylyl Imidodiphosphate - pharmacology ; Animals ; Beat frequencies ; Cell motility ; Chromatography ; Dyneins - metabolism ; Flagella ; Hydrolysis ; Male ; Microtubules ; Microtubules - metabolism ; Nucleotides ; Phosphatases ; Protein Binding ; Rapid Communications ; Sea Urchins ; Sperm Motility - drug effects ; Spermatozoa ; Spermatozoa - drug effects ; Spermatozoa - physiology</subject><ispartof>The Journal of cell biology, 1978-12, Vol.79 (3), p.827-832</ispartof><rights>Copyright 1978 The Rockefeller University Press</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3147-5076253ecbb4a5ed6e41573fbbe90578e46dff861189ba5d18b5348929778d723</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27923,27924</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/153347$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Penningroth, Stephen M.</creatorcontrib><creatorcontrib>Witman, George B.</creatorcontrib><title>Effects of Adenylyl Imidodiphosphate, a Nonhydrolyzable Adenosine Triphosphate Analog, on Reactivated and Rigor Wave Sea Urchin Sperm</title><title>The Journal of cell biology</title><addtitle>J Cell Biol</addtitle><description>A nonhydrolyzable ATP analog, adenylyl imidodiphosphate (AMP-PNP), has been used to study the role of ATP binding in flagellar motility. Sea urchin sperm of Lytechinus pictus were demembranated, reactivated, and locked in "rigor waves" by a modification of the method of Gibbons and Gibbons (11). Rigor wave sperm relaxed within 2 min after addition of 4 μM ATP, and reactivated upon addition of 10-12 μM ATP. The beat frequency of the reactivated sperm varied with ATP concentration according to Michaelis-Menten kinetics (`` Km''=0.24 mM;`` Vmax''=44 Hz) and was competitively inhibited by AMP-PNP (`` Ki''≅ 8.1 mM). Rigor wave sperm were completely relaxed (straightened) within 2 min by AMP-PNP at concentrations of 2-4 mM. The possibilities that relaxation in AMP-PNP was a result of ATP contamination, AMP-PNP hydrolysis, or lowering of the free Mg++concentration were conclusively ruled out. The results suggest that dynein cross-bridge release is dependent upon ATP binding but not hydrolysis.</description><subject>Adenosine triphosphatases</subject><subject>Adenosine Triphosphate - analogs & derivatives</subject><subject>Adenosine Triphosphate - metabolism</subject><subject>Adenosine Triphosphate - pharmacology</subject><subject>Adenylyl Imidodiphosphate - pharmacology</subject><subject>Animals</subject><subject>Beat frequencies</subject><subject>Cell motility</subject><subject>Chromatography</subject><subject>Dyneins - metabolism</subject><subject>Flagella</subject><subject>Hydrolysis</subject><subject>Male</subject><subject>Microtubules</subject><subject>Microtubules - metabolism</subject><subject>Nucleotides</subject><subject>Phosphatases</subject><subject>Protein Binding</subject><subject>Rapid Communications</subject><subject>Sea Urchins</subject><subject>Sperm Motility - drug effects</subject><subject>Spermatozoa</subject><subject>Spermatozoa - drug effects</subject><subject>Spermatozoa - physiology</subject><issn>0021-9525</issn><issn>1540-8140</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1978</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkc-L1DAUx4P4a1y9efSQk6fpmDRJk16EYVl1YVHYH3gMafM6zZAmNekM1Lv_t3V3cfT04H0_fN-DD0JvKdlQotiHfdtsZL1hG1XKJ2hFBSeFopw8RStCSlrUohQv0auc94QQLjl7gZ5TwRiXK_TrouugnTKOHd5aCLOfPb4cnI3WjX3MY28mWGODv8bQzzZFP_80jYd7OGYXAN-mE4m3wfi4W-MY8DWYdnLHZWuxCRZfu11M-Ls5Ar4Bg-9S27uAb0ZIw2v0rDM-w5vHeYbuPl3cnn8prr59vjzfXhUto1wWgsiqFAzapuFGgK2AUyFZ1zRQEyEV8Mp2naooVXVjhKWqEYyruqylVFaW7Ax9fOgdD80AtoUwJeP1mNxg0qyjcfr_JLhe7-JRl5SSUtKl4P1jQYo_DpAnPbjcgvcmQDxkLTmtS1KxBVw_gG2KOSfo_h6hRP-xphdrWtaa6cXagr_797ETfK_pFO_zFNMprYiqGGG_ARDQnpo</recordid><startdate>19781201</startdate><enddate>19781201</enddate><creator>Penningroth, Stephen M.</creator><creator>Witman, George B.</creator><general>Rockefeller University Press</general><general>The Rockefeller University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19781201</creationdate><title>Effects of Adenylyl Imidodiphosphate, a Nonhydrolyzable Adenosine Triphosphate Analog, on Reactivated and Rigor Wave Sea Urchin Sperm</title><author>Penningroth, Stephen M. ; Witman, George B.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3147-5076253ecbb4a5ed6e41573fbbe90578e46dff861189ba5d18b5348929778d723</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1978</creationdate><topic>Adenosine triphosphatases</topic><topic>Adenosine Triphosphate - analogs & derivatives</topic><topic>Adenosine Triphosphate - metabolism</topic><topic>Adenosine Triphosphate - pharmacology</topic><topic>Adenylyl Imidodiphosphate - pharmacology</topic><topic>Animals</topic><topic>Beat frequencies</topic><topic>Cell motility</topic><topic>Chromatography</topic><topic>Dyneins - metabolism</topic><topic>Flagella</topic><topic>Hydrolysis</topic><topic>Male</topic><topic>Microtubules</topic><topic>Microtubules - metabolism</topic><topic>Nucleotides</topic><topic>Phosphatases</topic><topic>Protein Binding</topic><topic>Rapid Communications</topic><topic>Sea Urchins</topic><topic>Sperm Motility - drug effects</topic><topic>Spermatozoa</topic><topic>Spermatozoa - drug effects</topic><topic>Spermatozoa - physiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Penningroth, Stephen M.</creatorcontrib><creatorcontrib>Witman, George B.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Penningroth, Stephen M.</au><au>Witman, George B.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Effects of Adenylyl Imidodiphosphate, a Nonhydrolyzable Adenosine Triphosphate Analog, on Reactivated and Rigor Wave Sea Urchin Sperm</atitle><jtitle>The Journal of cell biology</jtitle><addtitle>J Cell Biol</addtitle><date>1978-12-01</date><risdate>1978</risdate><volume>79</volume><issue>3</issue><spage>827</spage><epage>832</epage><pages>827-832</pages><issn>0021-9525</issn><eissn>1540-8140</eissn><abstract>A nonhydrolyzable ATP analog, adenylyl imidodiphosphate (AMP-PNP), has been used to study the role of ATP binding in flagellar motility. Sea urchin sperm of Lytechinus pictus were demembranated, reactivated, and locked in "rigor waves" by a modification of the method of Gibbons and Gibbons (11). Rigor wave sperm relaxed within 2 min after addition of 4 μM ATP, and reactivated upon addition of 10-12 μM ATP. The beat frequency of the reactivated sperm varied with ATP concentration according to Michaelis-Menten kinetics (`` Km''=0.24 mM;`` Vmax''=44 Hz) and was competitively inhibited by AMP-PNP (`` Ki''≅ 8.1 mM). Rigor wave sperm were completely relaxed (straightened) within 2 min by AMP-PNP at concentrations of 2-4 mM. The possibilities that relaxation in AMP-PNP was a result of ATP contamination, AMP-PNP hydrolysis, or lowering of the free Mg++concentration were conclusively ruled out. The results suggest that dynein cross-bridge release is dependent upon ATP binding but not hydrolysis.</abstract><cop>United States</cop><pub>Rockefeller University Press</pub><pmid>153347</pmid><doi>10.1083/jcb.79.3.827</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of cell biology, 1978-12, Vol.79 (3), p.827-832 |
| issn | 0021-9525 1540-8140 |
| language | eng |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Adenosine triphosphatases Adenosine Triphosphate - analogs & derivatives Adenosine Triphosphate - metabolism Adenosine Triphosphate - pharmacology Adenylyl Imidodiphosphate - pharmacology Animals Beat frequencies Cell motility Chromatography Dyneins - metabolism Flagella Hydrolysis Male Microtubules Microtubules - metabolism Nucleotides Phosphatases Protein Binding Rapid Communications Sea Urchins Sperm Motility - drug effects Spermatozoa Spermatozoa - drug effects Spermatozoa - physiology |
| title | Effects of Adenylyl Imidodiphosphate, a Nonhydrolyzable Adenosine Triphosphate Analog, on Reactivated and Rigor Wave Sea Urchin Sperm |
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