Alpha-isoenzyme of alcohol dehydrogenase from monkey liver. Cloning, expression, mechanism, coenzyme, and substrate specificity
The cDNA for the alpha-isoenzyme from rhesus monkey (Macaca mulatta) liver was cloned and expressed in yeast. The alpha-isoenzymes of human and monkey liver alcohol dehydrogenase differ from the other human and horse liver enzymes in having Met57, Ala93, and Val116 instead of Leu57, Phe93, and Leu11...
Gespeichert in:
| Veröffentlicht in: | The Journal of biological chemistry 1992-06, Vol.267 (18), p.12592-12599 |
|---|---|
| Hauptverfasser: | , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 12599 |
|---|---|
| container_issue | 18 |
| container_start_page | 12592 |
| container_title | The Journal of biological chemistry |
| container_volume | 267 |
| creator | LIGHT, D. R DENNIS, M. S FORSYTHE, I. J CHUNG-CHENG LIU GREEN, D. W KRATZER, D. A PLAPP, B. V |
| description | The cDNA for the alpha-isoenzyme from rhesus monkey (Macaca mulatta) liver was cloned and expressed in yeast. The alpha-isoenzymes
of human and monkey liver alcohol dehydrogenase differ from the other human and horse liver enzymes in having Met57, Ala93,
and Val116 instead of Leu57, Phe93, and Leu116 in the substrate binding pocket and Gly47 instead of Arg47 near the pyrophosphate
moiety of the coenzyme. The effects of these differences on the kinetic mechanism, substrate specificity, and coenzyme binding
were studied with the purified, recombinant monkey alpha-isoenzyme (MmADH alpha) and mutated enzymes with Gly47 substituted
with His or Arg. The mechanism appears to be random for the binding of NAD+ and ethanol and ordered for NADH and acetaldehyde,
with formation of a dead-end enzyme-NADH-ethanol complex. MmADH alpha reacts 130-fold slower (V/K) with ethanol and 3-25-fold
slower with 2-methyl alcohols but 20-fold faster with cyclohexanol, as compared with horse (Equus caballus) liver EE isoenzyme
(EqADH). MmADH alpha is stereoselective for the R isomer of 2-butanol, whereas EqADH favors the S isomer. Both enzymes have
comparable reactivity with larger primary alcohols. MmADH alpha is more reactive with secondary alcohols and has highest activity
with cyclohexanol. However, it does not react with steroids such as 5 beta-androstane-17 beta-ol-3-one. Molecular modeling
suggests that the differences between MmADH alpha and EqADH are a result of the substitution of Ala for Phe93 and Thr for
Ser48. MmADH alpha binds NAD+ most rapidly when a group with a pK of 7.4 is unprotonated, implicating His51 in this reaction.
The G47R substitution decreased the dissociation constants for NAD+ and NADH and turnover numbers only about 2-fold, whereas
the G47H substitution increased dissociation constants 7-14-fold and turnover numbers 4-fold. A basic residue at position
47 is not crucial for activity, as multiple interactions determine coenzyme affinity. |
| doi_str_mv | 10.1016/S0021-9258(18)42318-6 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_73046285</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>16517087</sourcerecordid><originalsourceid>FETCH-LOGICAL-c440t-1da9686e1fb88762815afebff35c34ba22560753c1547f635b007b59f68a843d3</originalsourceid><addsrcrecordid>eNqFkU9v1DAQxS0EKkvhI1TyASGQNsUT_4n3WK1aQKrEAZC4WY4z3hiSONhZIFz46mS7q_bIXOYwv_dmNI-QC2CXwEC9_cRYCcWmlPo16Dei5KAL9YisgGlecAlfH5PVPfKUPMv5G1tKbOCMnIECXSmxIn-vurG1RcgRhz9zjzR6ajsX29jRBtu5SXGHg81IfYo97ePwHWfahZ-YLum2i0MYdmuKv8eEOYc4rGmPrrVDyP2aupPpmtqhoXlf5ynZCWke0QUfXJjm5-SJt13GF6d-Tr7cXH_evi9uP777sL26LZwQbCqgsRulFYKv9XJ4qUFaj7X3XDoualuWUrFKcgdSVF5xWTNW1XLjlbZa8Iafk1dH3zHFH3vMk-lDdth1dsC4z6biTCy28r8gKAkV09UCyiPoUsw5oTdjCr1NswFmDgmZu4TM4f0GtLlLyKhFd3FasK97bB5Ux0iW-cvT3GZnO5_s4EK-x6SQwHj1gLVh1_4KCU0domuxN6WqDvuglJuS_wOrdqYO</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>16517087</pqid></control><display><type>article</type><title>Alpha-isoenzyme of alcohol dehydrogenase from monkey liver. Cloning, expression, mechanism, coenzyme, and substrate specificity</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Alma/SFX Local Collection</source><creator>LIGHT, D. R ; DENNIS, M. S ; FORSYTHE, I. J ; CHUNG-CHENG LIU ; GREEN, D. W ; KRATZER, D. A ; PLAPP, B. V</creator><creatorcontrib>LIGHT, D. R ; DENNIS, M. S ; FORSYTHE, I. J ; CHUNG-CHENG LIU ; GREEN, D. W ; KRATZER, D. A ; PLAPP, B. V</creatorcontrib><description>The cDNA for the alpha-isoenzyme from rhesus monkey (Macaca mulatta) liver was cloned and expressed in yeast. The alpha-isoenzymes
of human and monkey liver alcohol dehydrogenase differ from the other human and horse liver enzymes in having Met57, Ala93,
and Val116 instead of Leu57, Phe93, and Leu116 in the substrate binding pocket and Gly47 instead of Arg47 near the pyrophosphate
moiety of the coenzyme. The effects of these differences on the kinetic mechanism, substrate specificity, and coenzyme binding
were studied with the purified, recombinant monkey alpha-isoenzyme (MmADH alpha) and mutated enzymes with Gly47 substituted
with His or Arg. The mechanism appears to be random for the binding of NAD+ and ethanol and ordered for NADH and acetaldehyde,
with formation of a dead-end enzyme-NADH-ethanol complex. MmADH alpha reacts 130-fold slower (V/K) with ethanol and 3-25-fold
slower with 2-methyl alcohols but 20-fold faster with cyclohexanol, as compared with horse (Equus caballus) liver EE isoenzyme
(EqADH). MmADH alpha is stereoselective for the R isomer of 2-butanol, whereas EqADH favors the S isomer. Both enzymes have
comparable reactivity with larger primary alcohols. MmADH alpha is more reactive with secondary alcohols and has highest activity
with cyclohexanol. However, it does not react with steroids such as 5 beta-androstane-17 beta-ol-3-one. Molecular modeling
suggests that the differences between MmADH alpha and EqADH are a result of the substitution of Ala for Phe93 and Thr for
Ser48. MmADH alpha binds NAD+ most rapidly when a group with a pK of 7.4 is unprotonated, implicating His51 in this reaction.
The G47R substitution decreased the dissociation constants for NAD+ and NADH and turnover numbers only about 2-fold, whereas
the G47H substitution increased dissociation constants 7-14-fold and turnover numbers 4-fold. A basic residue at position
47 is not crucial for activity, as multiple interactions determine coenzyme affinity.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1016/S0021-9258(18)42318-6</identifier><identifier>PMID: 1618764</identifier><identifier>CODEN: JBCHA3</identifier><language>eng</language><publisher>Bethesda, MD: American Society for Biochemistry and Molecular Biology</publisher><subject>alcohol dehydrogenase ; Alcohol Dehydrogenase - chemistry ; Alcohol Dehydrogenase - genetics ; Alcohol Dehydrogenase - metabolism ; Analytical, structural and metabolic biochemistry ; Animals ; Base Sequence ; Binding Sites ; Biological and medical sciences ; cloning ; Cloning, Molecular ; Coenzymes - metabolism ; comparison ; Cyclohexanols - pharmacology ; Cyclohexanones - pharmacology ; DNA ; Enzymes and enzyme inhibitors ; expression ; Fundamental and applied biological sciences. Psychology ; Gene Expression ; Humans ; Hydrogen-Ion Concentration ; isoenzymes ; Isoenzymes - genetics ; Isoenzymes - metabolism ; Kinetics ; liver ; Liver - enzymology ; Macaca mulatta ; Models, Molecular ; Molecular Sequence Data ; molecular simulation ; Oxidoreductases ; Protein Conformation ; Substrate Specificity</subject><ispartof>The Journal of biological chemistry, 1992-06, Vol.267 (18), p.12592-12599</ispartof><rights>1992 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c440t-1da9686e1fb88762815afebff35c34ba22560753c1547f635b007b59f68a843d3</citedby><cites>FETCH-LOGICAL-c440t-1da9686e1fb88762815afebff35c34ba22560753c1547f635b007b59f68a843d3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=5451037$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1618764$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>LIGHT, D. R</creatorcontrib><creatorcontrib>DENNIS, M. S</creatorcontrib><creatorcontrib>FORSYTHE, I. J</creatorcontrib><creatorcontrib>CHUNG-CHENG LIU</creatorcontrib><creatorcontrib>GREEN, D. W</creatorcontrib><creatorcontrib>KRATZER, D. A</creatorcontrib><creatorcontrib>PLAPP, B. V</creatorcontrib><title>Alpha-isoenzyme of alcohol dehydrogenase from monkey liver. Cloning, expression, mechanism, coenzyme, and substrate specificity</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The cDNA for the alpha-isoenzyme from rhesus monkey (Macaca mulatta) liver was cloned and expressed in yeast. The alpha-isoenzymes
of human and monkey liver alcohol dehydrogenase differ from the other human and horse liver enzymes in having Met57, Ala93,
and Val116 instead of Leu57, Phe93, and Leu116 in the substrate binding pocket and Gly47 instead of Arg47 near the pyrophosphate
moiety of the coenzyme. The effects of these differences on the kinetic mechanism, substrate specificity, and coenzyme binding
were studied with the purified, recombinant monkey alpha-isoenzyme (MmADH alpha) and mutated enzymes with Gly47 substituted
with His or Arg. The mechanism appears to be random for the binding of NAD+ and ethanol and ordered for NADH and acetaldehyde,
with formation of a dead-end enzyme-NADH-ethanol complex. MmADH alpha reacts 130-fold slower (V/K) with ethanol and 3-25-fold
slower with 2-methyl alcohols but 20-fold faster with cyclohexanol, as compared with horse (Equus caballus) liver EE isoenzyme
(EqADH). MmADH alpha is stereoselective for the R isomer of 2-butanol, whereas EqADH favors the S isomer. Both enzymes have
comparable reactivity with larger primary alcohols. MmADH alpha is more reactive with secondary alcohols and has highest activity
with cyclohexanol. However, it does not react with steroids such as 5 beta-androstane-17 beta-ol-3-one. Molecular modeling
suggests that the differences between MmADH alpha and EqADH are a result of the substitution of Ala for Phe93 and Thr for
Ser48. MmADH alpha binds NAD+ most rapidly when a group with a pK of 7.4 is unprotonated, implicating His51 in this reaction.
The G47R substitution decreased the dissociation constants for NAD+ and NADH and turnover numbers only about 2-fold, whereas
the G47H substitution increased dissociation constants 7-14-fold and turnover numbers 4-fold. A basic residue at position
47 is not crucial for activity, as multiple interactions determine coenzyme affinity.</description><subject>alcohol dehydrogenase</subject><subject>Alcohol Dehydrogenase - chemistry</subject><subject>Alcohol Dehydrogenase - genetics</subject><subject>Alcohol Dehydrogenase - metabolism</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Binding Sites</subject><subject>Biological and medical sciences</subject><subject>cloning</subject><subject>Cloning, Molecular</subject><subject>Coenzymes - metabolism</subject><subject>comparison</subject><subject>Cyclohexanols - pharmacology</subject><subject>Cyclohexanones - pharmacology</subject><subject>DNA</subject><subject>Enzymes and enzyme inhibitors</subject><subject>expression</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression</subject><subject>Humans</subject><subject>Hydrogen-Ion Concentration</subject><subject>isoenzymes</subject><subject>Isoenzymes - genetics</subject><subject>Isoenzymes - metabolism</subject><subject>Kinetics</subject><subject>liver</subject><subject>Liver - enzymology</subject><subject>Macaca mulatta</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>molecular simulation</subject><subject>Oxidoreductases</subject><subject>Protein Conformation</subject><subject>Substrate Specificity</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU9v1DAQxS0EKkvhI1TyASGQNsUT_4n3WK1aQKrEAZC4WY4z3hiSONhZIFz46mS7q_bIXOYwv_dmNI-QC2CXwEC9_cRYCcWmlPo16Dei5KAL9YisgGlecAlfH5PVPfKUPMv5G1tKbOCMnIECXSmxIn-vurG1RcgRhz9zjzR6ajsX29jRBtu5SXGHg81IfYo97ePwHWfahZ-YLum2i0MYdmuKv8eEOYc4rGmPrrVDyP2aupPpmtqhoXlf5ynZCWke0QUfXJjm5-SJt13GF6d-Tr7cXH_evi9uP777sL26LZwQbCqgsRulFYKv9XJ4qUFaj7X3XDoualuWUrFKcgdSVF5xWTNW1XLjlbZa8Iafk1dH3zHFH3vMk-lDdth1dsC4z6biTCy28r8gKAkV09UCyiPoUsw5oTdjCr1NswFmDgmZu4TM4f0GtLlLyKhFd3FasK97bB5Ux0iW-cvT3GZnO5_s4EK-x6SQwHj1gLVh1_4KCU0domuxN6WqDvuglJuS_wOrdqYO</recordid><startdate>19920625</startdate><enddate>19920625</enddate><creator>LIGHT, D. R</creator><creator>DENNIS, M. S</creator><creator>FORSYTHE, I. J</creator><creator>CHUNG-CHENG LIU</creator><creator>GREEN, D. W</creator><creator>KRATZER, D. A</creator><creator>PLAPP, B. V</creator><general>American Society for Biochemistry and Molecular Biology</general><scope>IQODW</scope><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>7QL</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M81</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19920625</creationdate><title>Alpha-isoenzyme of alcohol dehydrogenase from monkey liver. Cloning, expression, mechanism, coenzyme, and substrate specificity</title><author>LIGHT, D. R ; DENNIS, M. S ; FORSYTHE, I. J ; CHUNG-CHENG LIU ; GREEN, D. W ; KRATZER, D. A ; PLAPP, B. V</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c440t-1da9686e1fb88762815afebff35c34ba22560753c1547f635b007b59f68a843d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>alcohol dehydrogenase</topic><topic>Alcohol Dehydrogenase - chemistry</topic><topic>Alcohol Dehydrogenase - genetics</topic><topic>Alcohol Dehydrogenase - metabolism</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Binding Sites</topic><topic>Biological and medical sciences</topic><topic>cloning</topic><topic>Cloning, Molecular</topic><topic>Coenzymes - metabolism</topic><topic>comparison</topic><topic>Cyclohexanols - pharmacology</topic><topic>Cyclohexanones - pharmacology</topic><topic>DNA</topic><topic>Enzymes and enzyme inhibitors</topic><topic>expression</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression</topic><topic>Humans</topic><topic>Hydrogen-Ion Concentration</topic><topic>isoenzymes</topic><topic>Isoenzymes - genetics</topic><topic>Isoenzymes - metabolism</topic><topic>Kinetics</topic><topic>liver</topic><topic>Liver - enzymology</topic><topic>Macaca mulatta</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>molecular simulation</topic><topic>Oxidoreductases</topic><topic>Protein Conformation</topic><topic>Substrate Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>LIGHT, D. R</creatorcontrib><creatorcontrib>DENNIS, M. S</creatorcontrib><creatorcontrib>FORSYTHE, I. J</creatorcontrib><creatorcontrib>CHUNG-CHENG LIU</creatorcontrib><creatorcontrib>GREEN, D. W</creatorcontrib><creatorcontrib>KRATZER, D. A</creatorcontrib><creatorcontrib>PLAPP, B. V</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biochemistry Abstracts 3</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>LIGHT, D. R</au><au>DENNIS, M. S</au><au>FORSYTHE, I. J</au><au>CHUNG-CHENG LIU</au><au>GREEN, D. W</au><au>KRATZER, D. A</au><au>PLAPP, B. V</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Alpha-isoenzyme of alcohol dehydrogenase from monkey liver. Cloning, expression, mechanism, coenzyme, and substrate specificity</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1992-06-25</date><risdate>1992</risdate><volume>267</volume><issue>18</issue><spage>12592</spage><epage>12599</epage><pages>12592-12599</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><coden>JBCHA3</coden><abstract>The cDNA for the alpha-isoenzyme from rhesus monkey (Macaca mulatta) liver was cloned and expressed in yeast. The alpha-isoenzymes
of human and monkey liver alcohol dehydrogenase differ from the other human and horse liver enzymes in having Met57, Ala93,
and Val116 instead of Leu57, Phe93, and Leu116 in the substrate binding pocket and Gly47 instead of Arg47 near the pyrophosphate
moiety of the coenzyme. The effects of these differences on the kinetic mechanism, substrate specificity, and coenzyme binding
were studied with the purified, recombinant monkey alpha-isoenzyme (MmADH alpha) and mutated enzymes with Gly47 substituted
with His or Arg. The mechanism appears to be random for the binding of NAD+ and ethanol and ordered for NADH and acetaldehyde,
with formation of a dead-end enzyme-NADH-ethanol complex. MmADH alpha reacts 130-fold slower (V/K) with ethanol and 3-25-fold
slower with 2-methyl alcohols but 20-fold faster with cyclohexanol, as compared with horse (Equus caballus) liver EE isoenzyme
(EqADH). MmADH alpha is stereoselective for the R isomer of 2-butanol, whereas EqADH favors the S isomer. Both enzymes have
comparable reactivity with larger primary alcohols. MmADH alpha is more reactive with secondary alcohols and has highest activity
with cyclohexanol. However, it does not react with steroids such as 5 beta-androstane-17 beta-ol-3-one. Molecular modeling
suggests that the differences between MmADH alpha and EqADH are a result of the substitution of Ala for Phe93 and Thr for
Ser48. MmADH alpha binds NAD+ most rapidly when a group with a pK of 7.4 is unprotonated, implicating His51 in this reaction.
The G47R substitution decreased the dissociation constants for NAD+ and NADH and turnover numbers only about 2-fold, whereas
the G47H substitution increased dissociation constants 7-14-fold and turnover numbers 4-fold. A basic residue at position
47 is not crucial for activity, as multiple interactions determine coenzyme affinity.</abstract><cop>Bethesda, MD</cop><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>1618764</pmid><doi>10.1016/S0021-9258(18)42318-6</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9258 |
| ispartof | The Journal of biological chemistry, 1992-06, Vol.267 (18), p.12592-12599 |
| issn | 0021-9258 1083-351X |
| language | eng |
| recordid | cdi_proquest_miscellaneous_73046285 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | alcohol dehydrogenase Alcohol Dehydrogenase - chemistry Alcohol Dehydrogenase - genetics Alcohol Dehydrogenase - metabolism Analytical, structural and metabolic biochemistry Animals Base Sequence Binding Sites Biological and medical sciences cloning Cloning, Molecular Coenzymes - metabolism comparison Cyclohexanols - pharmacology Cyclohexanones - pharmacology DNA Enzymes and enzyme inhibitors expression Fundamental and applied biological sciences. Psychology Gene Expression Humans Hydrogen-Ion Concentration isoenzymes Isoenzymes - genetics Isoenzymes - metabolism Kinetics liver Liver - enzymology Macaca mulatta Models, Molecular Molecular Sequence Data molecular simulation Oxidoreductases Protein Conformation Substrate Specificity |
| title | Alpha-isoenzyme of alcohol dehydrogenase from monkey liver. Cloning, expression, mechanism, coenzyme, and substrate specificity |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-28T17%3A51%3A43IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Alpha-isoenzyme%20of%20alcohol%20dehydrogenase%20from%20monkey%20liver.%20Cloning,%20expression,%20mechanism,%20coenzyme,%20and%20substrate%20specificity&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=LIGHT,%20D.%20R&rft.date=1992-06-25&rft.volume=267&rft.issue=18&rft.spage=12592&rft.epage=12599&rft.pages=12592-12599&rft.issn=0021-9258&rft.eissn=1083-351X&rft.coden=JBCHA3&rft_id=info:doi/10.1016/S0021-9258(18)42318-6&rft_dat=%3Cproquest_cross%3E16517087%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=16517087&rft_id=info:pmid/1618764&rfr_iscdi=true |