;PMID: 2848026 ;source_file_1804.mrg ;Release 0.9 of PennBioIE ;Funded by NSF ITR EIA-0205448 ;0)section:[e:0..41] = [t:0..41] ;1)sentence:[e:47..184] = [t:47..184] ;2)section:[e:188..218] = [t:188..218] ;3)section:[e:222..298] = [t:222..298] ;4)sentence:[e:302..563] = [t:302..563] ;5)sentence:[e:564..697] = [t:564..697] ;6)sentence:[e:698..814] = [t:698..814] ;7)sentence:[e:815..940] = [t:815..940] ;8)sentence:[e:941..1142] = [t:941..1142] ;9)sentence:[e:1143..1429] = [t:1143..1429] ;10)sentence:[e:1430..1708] = [t:1430..1708] ;11)sentence:[e:1709..1833] = [t:1709..1833] ;12)sentence:[e:1834..2007] = [t:1834..2007] ;13)sentence:[e:2008..2119] = [t:2008..2119] ;14)sentence:[e:2120..2210] = [t:2120..2210] ;15)sentence:[e:2211..2292] = [t:2211..2292] ;16)section:[e:2296..2340] = [t:2296..2340] ;section 0 Span:0..41 ;J Biol Chem. 1988 Dec 5;263(34):18138-42. (SEC (FRAG (NNP:[0..1] J) (NNP:[2..6] Biol) (NNP:[7..11] Chem) (.:[11..12] .) (CD:[13..17] 1988) (NNP:[18..21] Dec) (CC:[22..28] 5;263-LRB-) (CD:[28..30] 34) (-RRB-:[30..31] -RRB-) (::[31..32] :) (CD:[32..40] 18138-42) (.:[40..41] .))) ;sentence 1 Span:47..184 ;Purification and characterization of 5 beta-cholestane-3 alpha,7 alpha,12 ;alpha-triol 27-hydroxylase from female rat liver mitochondria. ;[84..148]:cyp450:"5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol ;27-hydroxylase" (SENT (NP-HLN (NP (NN:[47..59] Purification) (CC:[60..63] and) (NN:[64..80] characterization)) (PP (IN:[81..83] of) (NP (NP (CD:[84..85] 5) (NN:[86..103] beta-cholestane-3) (NN:[104..111] alpha,7) (NN:[112..120] alpha,12) (NN:[122..133] alpha-triol) (NN:[134..148] 27-hydroxylase)) (PP (IN:[149..153] from) (NP (JJ:[154..160] female) (NN:[161..164] rat) (NN:[165..170] liver) (NNS:[171..183] mitochondria))))) (.:[183..184] .))) ;section 2 Span:188..218 ;Okuda K, Masumoto O, Ohyama Y. (SEC (FRAG (NNP:[188..193] Okuda) (NNP:[194..196] K,) (NNP:[197..205] Masumoto) (NNP:[206..207] O) (,:[207..208] ,) (NNP:[209..215] Ohyama) (NNP:[216..217] Y) (.:[217..218] .))) ;section 3 Span:222..298 ;Department of Biochemistry, Hiroshima University School of Dentistry, Japan. (SEC (FRAG (NNP:[222..232] Department) (IN:[233..235] of) (NNP:[236..248] Biochemistry) (,:[248..249] ,) (NNP:[250..259] Hiroshima) (NNP:[260..270] University) (NNP:[271..277] School) (IN:[278..280] of) (NNP:[281..290] Dentistry) (,:[290..291] ,) (NNP:[292..297] Japan) (.:[297..298] .))) ;sentence 4 Span:302..563 ;5 beta-Cholestane-3 alpha,7 alpha,12 alpha-triol 27-hydroxylase (5 ;beta-cholestane-3 alpha,7 alpha,12 alpha-triol, NADPH:oxygen oxidoreductase ;(26-hydroxylating), EC 1.14.13.15) was purified from female rat liver ;mitochondria based on its catalytic activity. ;[302..365]:cyp450:"5 beta-Cholestane-3 alpha,7 alpha,12 alpha-triol ;27-hydroxylase" ;[367..416]:substance:"5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol" ;[418..445]:substance:"NADPH:oxygen oxidoreductase" (SENT (S (NP-SBJ-1 (NP (CD:[302..303] 5) (NN:[304..321] beta-Cholestane-3) (NN:[322..329] alpha,7) (NN:[330..338] alpha,12) (NN:[339..350] alpha-triol) (NN:[351..365] 27-hydroxylase)) (NP (-LRB-:[366..367] -LRB-) (NP (CD:[367..368] 5) (NN:[370..387] beta-cholestane-3) (NN:[388..395] alpha,7) (NN:[396..404] alpha,12) (NN:[405..416] alpha-triol)) (,:[416..417] ,) (NP (NP (NML (NN:[418..423] NADPH) (SYM:[423..424] :) (NN:[424..430] oxygen)) (NN:[431..445] oxidoreductase)) (NP (-LRB-:[447..448] -LRB-) (VBG:[448..464] 26-hydroxylating) (-RRB-:[464..465] -RRB-))) (,:[465..466] ,) (NP (NNP:[467..469] EC) (CD:[470..480] 1.14.13.15)) (-RRB-:[480..481] -RRB-))) (VP (VBD:[482..485] was) (VP (VBN:[486..494] purified) (NP-1 (-NONE-:[494..494] *)) (PP (IN:[495..499] from) (NP (JJ:[500..506] female) (NN:[507..510] rat) (NN:[511..516] liver) (NNS:[518..530] mitochondria))) (S-ADV (NP-SBJ (-NONE-:[530..530] *)) (VP (VBN:[531..536] based) (NP (-NONE-:[536..536] *)) (PP-CLR (IN:[537..539] on) (NP (PRP$:[540..543] its) (JJ:[544..553] catalytic) (NN:[554..562] activity))))))) (.:[562..563] .))) ;sentence 5 Span:564..697 ;The final preparation of the enzyme showed a single major band on the sodium ;dodecyl sulfate-polyacrylamide gel electrophoretogram. ;[594..600]:substance:"enzyme" ;[635..657]:substance:"sodium dodecyl sulfate" ;[658..672]:substance:"polyacrylamide" (SENT (S (NP-SBJ (NP (DT:[564..567] The) (JJ:[568..573] final) (NN:[574..585] preparation)) (PP (IN:[586..588] of) (NP (DT:[589..592] the) (NN:[594..600] enzyme)))) (VP (VBD:[601..607] showed) (NP (DT:[608..609] a) (JJ:[610..616] single) (JJ:[617..622] major) (NN:[623..627] band)) (PP-LOC (IN:[628..630] on) (NP (DT:[631..634] the) (NML (NML (NML (NN:[635..641] sodium) (NN:[642..649] dodecyl) (NN:[650..657] sulfate)) (HYPH:[657..658] -) (NN:[658..672] polyacrylamide)) (NN:[674..677] gel)) (NN:[678..696] electrophoretogram)))) (.:[696..697] .))) ;sentence 6 Span:698..814 ;The content of purified enzyme was 12 nmol/mg of protein, and the specific ;activity was 431 nmol/min/mg of protein. ;[722..728]:substance:"enzyme" ;[733..735]:quantitative-value:"12" ;[736..755]:quantitative-units:"nmol/mg of protein" ;[787..790]:quantitative-value:"431" ;[791..813]:quantitative-units:"nmol/min/mg of protein" (SENT (S (S (NP-SBJ (NP (DT:[698..701] The) (NN:[702..709] content)) (PP (IN:[710..712] of) (NP (VBN:[713..721] purified) (NN:[722..728] enzyme)))) (VP (VBD:[729..732] was) (NP-PRD (NP (CD:[733..735] 12) (NN:[736..740] nmol)) (PP (SYM:[740..741] /) (NP (NP (NN:[741..743] mg)) (PP (IN:[744..746] of) (NP (NN:[748..755] protein)))))))) (,:[755..756] ,) (CC:[757..760] and) (S (NP-SBJ (DT:[761..764] the) (JJ:[765..773] specific) (NN:[774..782] activity)) (VP (VBD:[783..786] was) (NP-PRD (NP (CD:[787..790] 431) (NN:[791..795] nmol)) (PP (SYM:[795..796] /) (NP (NN:[796..799] min))) (PP (SYM:[799..800] /) (NP (NP (NN:[800..802] mg)) (PP (IN:[803..805] of) (NP (NN:[806..813] protein)))))))) (.:[813..814] .))) ;sentence 7 Span:815..940 ;The molecular weight of the enzyme was determined from sodium dodecyl ;sulfate-polyacrylamide gel electrophoresis as 52,500. ;[844..850]:substance:"enzyme" ;[871..893]:substance:"sodium dodecyl sulfate" ;[894..908]:substance:"polyacrylamide" ;[933..939]:quantitative-value:"52,500" (SENT (S (NP-SBJ-1 (NP (DT:[815..818] The) (JJ:[819..828] molecular) (NN:[830..836] weight)) (PP (IN:[837..839] of) (NP (DT:[840..843] the) (NN:[844..850] enzyme)))) (VP (VBD:[851..854] was) (VP (VBN:[855..865] determined) (NP-1 (-NONE-:[865..865] *)) (PP (IN:[866..870] from) (NP (NML (NML (NML (NN:[871..877] sodium) (NN:[878..885] dodecyl) (NN:[886..893] sulfate)) (HYPH:[893..894] -) (NN:[894..908] polyacrylamide)) (NN:[910..913] gel)) (NN:[914..929] electrophoresis))) (PP (IN:[930..932] as) (NP (CD:[933..939] 52,500))))) (.:[939..940] .))) ;sentence 8 Span:941..1142 ;The absorption spectra of the purified enzyme and that of the ;dithionite-reduced CO complex showed peaks at 417 and 450 nm, respectively, ;indicating the enzyme belongs to the cytochrome P-450 family. ;[980..986]:cyp450:"enzyme" ;[1004..1014]:substance:"dithionite" ;[1023..1033]:substance:"CO complex" ;[1050..1053]:quantitative-value:"417" ;[1058..1061]:quantitative-value:"450" ;[1062..1064]:quantitative-units:"nm" ;[1096..1102]:cyp450:"enzyme" ;[1118..1134]:cyp450:"cytochrome P-450" (SENT (S (NP-SBJ (NP (NP (DT:[941..944] The) (NN:[945..955] absorption) (NNS:[956..963] spectra)) (PP (IN:[964..966] of) (NP (DT:[967..970] the) (VBN:[971..979] purified) (NN:[980..986] enzyme)))) (CC:[987..990] and) (NP (NP (DT:[992..996] that)) (PP (IN:[997..999] of) (NP (DT:[1000..1003] the) (ADJP (NN:[1004..1014] dithionite) (HYPH:[1014..1015] -) (VBN:[1015..1022] reduced)) (NN:[1023..1025] CO) (NN:[1026..1033] complex))))) (VP (VBD:[1034..1040] showed) (NP (NNS:[1041..1046] peaks)) (PP-LOC (IN:[1047..1049] at) (NP (NP (CD:[1050..1053] 417) (NML-1 (-NONE-:[1053..1053] *P*))) (CC:[1054..1057] and) (NP (CD:[1058..1061] 450) (NML-1 (NN:[1062..1064] nm))))) (,:[1064..1065] ,) (ADVP (RB:[1067..1079] respectively)) (,:[1079..1080] ,) (S-MNR (NP-SBJ (-NONE-:[1080..1080] *)) (VP (VBG:[1081..1091] indicating) (SBAR (-NONE-:[1091..1091] 0) (S (NP-SBJ (DT:[1092..1095] the) (NN:[1096..1102] enzyme)) (VP (VBZ:[1103..1110] belongs) (PP-CLR (TO:[1111..1113] to) (NP (DT:[1114..1117] the) (NML (NN:[1118..1128] cytochrome) (NN:[1129..1134] P-450)) (NN:[1135..1141] family))))))))) (.:[1141..1142] .))) ;sentence 9 Span:1143..1429 ;Upon reconstitution with the electron-transferring system of the adrenal ;(adrenodoxin and NADPH-adrenodoxin reductase), the enzyme showed high ;activity hydroxylating 5 beta-cholestane-3 alpha,7 alpha-12-triol at ;position 27 with a turnover number of 35.5 min-1 and Km of 6.3 microM. ;[1218..1229]:substance:"adrenodoxin" ;[1235..1262]:substance:"NADPH-adrenodoxin reductase" ;[1269..1275]:substance:"enzyme" ;[1312..1354]:substance:"5 beta-cholestane-3 alpha,7 alpha-12-triol" ;[1397..1401]:quantitative-value:"35.5" ;[1402..1407]:quantitative-units:"min-1" ;[1412..1414]:quantitative-name:"Km" ;[1418..1421]:quantitative-value:"6.3" ;[1422..1428]:quantitative-units:"microM" (SENT (S (PP-TMP (IN:[1143..1147] Upon) (NP (NP (NN:[1149..1163] reconstitution)) (PP (IN:[1164..1168] with) (NP (NP (DT:[1169..1172] the) (ADJP (NN:[1173..1181] electron) (HYPH:[1181..1182] -) (VBG:[1182..1194] transferring)) (NN:[1195..1201] system)) (PP (IN:[1202..1204] of) (NP (DT:[1205..1208] the) (JJ:[1209..1216] adrenal) (PRN (-LRB-:[1217..1218] -LRB-) (NP (NP (NN:[1218..1229] adrenodoxin)) (CC:[1231..1234] and) (NP (NML (NN:[1235..1240] NADPH) (HYPH:[1240..1241] -) (NN:[1241..1252] adrenodoxin)) (NN:[1253..1262] reductase))) (-RRB-:[1262..1263] -RRB-)))))))) (,:[1263..1264] ,) (NP-SBJ (DT:[1265..1268] the) (NN:[1269..1275] enzyme)) (VP (VBD:[1276..1282] showed) (NP (NP (JJ:[1283..1287] high) (NN:[1288..1296] activity)) (VP (VBG:[1297..1310] hydroxylating) (NP (CD:[1312..1313] 5) (NN:[1314..1331] beta-cholestane-3) (NN:[1332..1339] alpha,7) (NN:[1340..1354] alpha-12-triol)) (PP-LOC (IN:[1355..1357] at) (NP (NN:[1358..1366] position) (CD:[1367..1369] 27))) (PP (IN:[1370..1374] with) (NP (NP (NP (DT:[1375..1376] a) (NN:[1377..1385] turnover) (NN:[1386..1392] number)) (PP (IN:[1394..1396] of) (NP (CD:[1397..1401] 35.5) (NN:[1402..1407] min-1)))) (CC:[1408..1411] and) (NP (NP (NN:[1412..1414] Km)) (PP (IN:[1415..1417] of) (NP (CD:[1418..1421] 6.3) (NN:[1422..1428] microM))))))))) (.:[1428..1429] .))) ;sentence 10 Span:1430..1708 ;The enzyme activity was completely lost when the electron-transferring ;system was replaced by that of microsomes (NADPH-cytochrome P-450 reductase ;purified from rat liver microsomes), confirming that the P-450 enzyme was of ;the mitochondrial type, but not of the microsomal. ;[1434..1440]:substance:"enzyme" ;[1546..1578]:substance:"NADPH-cytochrome P-450 reductase" ;[1637..1649]:cyp450:"P-450 enzyme" (SENT (S (NP-SBJ-5 (DT:[1430..1433] The) (NN:[1434..1440] enzyme) (NN:[1441..1449] activity)) (VP (VBD:[1450..1453] was) (ADVP-EXT (RB:[1454..1464] completely)) (VP (VBN:[1465..1469] lost) (NP-5 (-NONE-:[1469..1469] *)) (SBAR-ADV (WHADVP-4 (WRB:[1470..1474] when)) (S (NP-SBJ-3 (DT:[1476..1479] the) (ADJP (NN:[1480..1488] electron) (HYPH:[1488..1489] -) (VBG:[1489..1501] transferring)) (NN:[1502..1508] system)) (VP (VBD:[1509..1512] was) (VP (VBN:[1513..1521] replaced) (NP-3 (-NONE-:[1521..1521] *)) (PP (IN:[1522..1524] by) (NP-LGS (NP (DT:[1525..1529] that)) (PP (IN:[1530..1532] of) (NP (NNS:[1533..1543] microsomes) (PRN (-LRB-:[1545..1546] -LRB-) (NP (NP (NML (NML (NN:[1546..1551] NADPH)) (HYPH:[1551..1552] -) (NML (NN:[1552..1562] cytochrome) (NN:[1563..1568] P-450))) (NN:[1569..1578] reductase)) (VP (VBN:[1579..1587] purified) (NP (-NONE-:[1587..1587] *)) (PP (IN:[1588..1592] from) (NP (NN:[1593..1596] rat) (NN:[1597..1602] liver) (NNS:[1603..1613] microsomes))))) (-RRB-:[1613..1614] -RRB-)))))) (,:[1614..1615] ,) (S-ADV (NP-SBJ (-NONE-:[1615..1615] *)) (VP (VBG:[1617..1627] confirming) (SBAR (IN:[1628..1632] that) (S (NP-SBJ (DT:[1633..1636] the) (NN:[1637..1642] P-450) (NN:[1643..1649] enzyme)) (VP (VP (VBD:[1650..1653] was) (PP-PRD (IN:[1654..1656] of) (NP=1 (DT:[1657..1660] the) (JJ:[1661..1674] mitochondrial) (NN:[1675..1679] type)))) (,:[1679..1680] ,) (CC:[1681..1684] but) (VP (RB:[1685..1688] not) (PP-PRD (IN:[1689..1691] of) (NP=1 (DT:[1692..1695] the) (JJ:[1697..1707] microsomal))))))))))))))) (.:[1707..1708] .))) ;sentence 11 Span:1709..1833 ;The omission of cytochrome P-450, adrenodoxin, or NADPH-adrenodoxin ;reductase resulted in complete loss of enzyme activity. ;[1725..1741]:cyp450:"cytochrome P-450" ;[1743..1754]:substance:"adrenodoxin" ;[1759..1787]:substance:"NADPH-adrenodoxin reductase" ;[1817..1823]:substance:"enzyme" (SENT (S (NP-SBJ (NP (DT:[1709..1712] The) (NN:[1713..1721] omission)) (PP (IN:[1722..1724] of) (NP (NP (NN:[1725..1735] cytochrome) (NN:[1736..1741] P-450)) (,:[1741..1742] ,) (NP (NN:[1743..1754] adrenodoxin)) (,:[1754..1755] ,) (CC:[1756..1758] or) (NP (NML (NN:[1759..1764] NADPH) (HYPH:[1764..1765] -) (NN:[1765..1776] adrenodoxin)) (NN:[1778..1787] reductase))))) (VP (VBD:[1788..1796] resulted) (PP-CLR (IN:[1797..1799] in) (NP (NP (JJ:[1800..1808] complete) (NN:[1809..1813] loss)) (PP (IN:[1814..1816] of) (NP (NN:[1817..1823] enzyme) (NN:[1824..1832] activity)))))) (.:[1832..1833] .))) ;sentence 12 Span:1834..2007 ;The specific activity toward 5 beta-cholestane-3 alpha, 7 alpha-diol was ;less than one-half that toward cholestanetriol and that toward cholesterol ;was about one-fiftieth. ;[1864..1903]:substance:"5 beta-cholestane-3 alpha, 7 alpha-diol" ;[1940..1955]:substance:"cholestanetriol" ;[1972..1983]:substance:"cholesterol" (SENT (S (S (NP-SBJ (NP (DT:[1834..1837] The) (JJ:[1838..1846] specific) (NN:[1847..1855] activity)) (PP (IN:[1857..1863] toward) (NP (CD:[1864..1865] 5) (NN:[1866..1883] beta-cholestane-3) (SYM:[1884..1889] alpha) (,:[1889..1890] ,) (CD:[1891..1892] 7) (NN:[1893..1903] alpha-diol)))) (VP (VBD:[1904..1907] was) (NP-PRD (NP (QP (JJR:[1908..1912] less) (IN:[1913..1917] than) (CD:[1918..1921] one) (HYPH:[1921..1922] -) (NN:[1922..1926] half)) (DT:[1927..1931] that)) (PP (IN:[1933..1939] toward) (NP (NN:[1940..1955] cholestanetriol)))))) (CC:[1956..1959] and) (S (NP-SBJ (NP (DT:[1960..1964] that)) (PP (IN:[1965..1971] toward) (NP (NN:[1972..1983] cholesterol)))) (VP (VBD:[1984..1987] was) (NP-PRD (QP (RB:[1988..1993] about) (CD:[1994..1997] one) (HYPH:[1997..1998] -) (NN:[1998..2006] fiftieth))))) (.:[2006..2007] .))) ;sentence 13 Span:2008..2119 ;The enzyme showed no activity toward xenobiotics such as benzphetamine, ;7-ethoxycoumarin, and benzo[a]pyrene. ;[2013..2019]:substance:"enzyme" ;[2046..2057]:substance:"xenobiotics" ;[2066..2079]:substance:"benzphetamine" ;[2082..2098]:substance:"7-ethoxycoumarin" ;[2104..2118]:substance:"benzo[a]pyrene" (SENT (S (NP-SBJ (DT:[2008..2011] The) (NN:[2013..2019] enzyme)) (VP (VBD:[2020..2026] showed) (NP (NP (DT:[2027..2029] no) (NN:[2030..2038] activity)) (PP (IN:[2039..2045] toward) (NP (NP (NNS:[2046..2057] xenobiotics)) (PP (JJ:[2058..2062] such) (IN:[2063..2065] as) (NP (NN:[2066..2079] benzphetamine) (,:[2079..2080] ,) (NN:[2082..2098] 7-ethoxycoumarin) (,:[2098..2099] ,) (CC:[2100..2103] and) (NN:[2104..2118] benzo-LSB-a-RSB-pyrene))))))) (.:[2118..2119] .))) ;sentence 14 Span:2120..2210 ;Its activity was not inhibited by metyrapone and slightly inhibited by ;aminoglutethimide. ;[2155..2165]:substance:"metyrapone" ;[2192..2209]:substance:"aminoglutethimide" (SENT (S (NP-SBJ-1 (PRP$:[2120..2123] Its) (NN:[2124..2132] activity)) (VP (VBD:[2133..2136] was) (VP (VP (RB:[2137..2140] not) (VBN:[2141..2150] inhibited) (NP-1 (-NONE-:[2150..2150] *)) (PP (IN:[2151..2153] by) (NP-LGS (NN:[2155..2165] metyrapone)))) (CC:[2166..2169] and) (VP (ADVP-EXT (RB:[2170..2178] slightly)) (VBN:[2179..2188] inhibited) (NP-1 (-NONE-:[2188..2188] *)) (PP (IN:[2189..2191] by) (NP-LGS (NN:[2192..2209] aminoglutethimide)))))) (.:[2209..2210] .))) ;sentence 15 Span:2211..2292 ;The enzyme activity was markedly lowered in an atmosphere of CO/O2/N2, ;40/20/40. ;[2215..2221]:substance:"enzyme" ;[2273..2275]:substance:"CO" ;[2276..2278]:substance:"O2" ;[2279..2281]:substance:"N2" (SENT (S (NP-SBJ-1 (DT:[2211..2214] The) (NN:[2215..2221] enzyme) (NN:[2222..2230] activity)) (VP (VBD:[2231..2234] was) (ADVP-MNR (RB:[2236..2244] markedly)) (VP (VBN:[2245..2252] lowered) (NP-1 (-NONE-:[2252..2252] *)) (PP-LOC (IN:[2253..2255] in) (NP (NP (DT:[2256..2258] an) (NN:[2259..2269] atmosphere)) (PP (IN:[2270..2272] of) (NP (NP (NN:[2273..2275] CO) (SYM:[2275..2276] /) (NN:[2276..2278] O2) (SYM:[2278..2279] /) (NN:[2279..2281] N2)) (,:[2281..2282] ,) (NP (CD:[2283..2285] 40) (SYM:[2285..2286] /) (CD:[2286..2288] 20) (SYM:[2288..2289] /) (CD:[2289..2291] 40)))))))) (.:[2291..2292] .))) ;section 16 Span:2296..2340 ;PMID: 2848026 [PubMed - indexed for MEDLINE] (SEC (FRAG (NNP:[2296..2300] PMID) (::[2300..2301] :) (CD:[2302..2309] 2848026) (NN:[2310..2311] -LSB-) (NNP:[2311..2317] PubMed) (::[2318..2319] -) (NN:[2320..2327] indexed) (IN:[2328..2331] for) (NNP:[2332..2340] MEDLINE-RSB-)))