By Michael Barany
This priceless source offers a scientific account of the biochemistry of soft muscle contraction. As a finished consultant to this quickly transforming into region of study, it covers the constitution and attribute homes of contractile and regulatory proteins, with unique emphasis on their estimated functionality within the stay muscle. additionally incorporated during this booklet are intermediate filament proteins, and desmin and vimentin, whose functionality in gentle muscle is unknown; and a number of other enzymes occupied with the phosphorylation-dephosphorylation of contractile and different proteins
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Additional info for Biochemistry of Smooth Muscle Contraction
Sample text
Note that, except for avian gizzard HC204, all isoforms of smooth muscle HC204 contain a serine residue (marked for rat aorta with an asterisk), which can be phosphorylated by casein kinase II in vitro (see Section II). This sequence is not found in the HC200 isoform. Although this site has also been found to be phosphorylated in intact cells, the function of the phosphorylation is not presently known (Kelley and Adelstein, 1990). 2. , 1993). , 1992). 69:1 in both human adult and infant bronchial tissue.
5 mM ATP ([~/-32p]ATP, 192 Ci/mol) for 30 min (Kelley and Adelstein, 1990). 6 mol of phosphate can be incorporated into the single residue phosphorylated, provided the myosin is first dephosphorylated. Techniques similar to those described by B~r~iny and B~r~ny in Chapter 2, this volume, but substituting buffers that are appropriate for chromatography of HC peptides, were used to map the single tryptic peptide generated following phosphorylation of HC204. C. Preparation of Antibodies to the 204- and 200-kDa Heavy Chains Polyclonal antibodies that detect both the 204- and 200-kDa HCs were raised in rabbits using purified bovine aortic smooth muscle myosin, which is a 1:1 mixture of both isoforms as antigens.
In general, the other actin binding sites that are postulated to be more stereospecific are considerably more conserved between 10 ROBERT S. A D E L S T E I N A N D J A M E S R. S E L L E R S ,,,L~17 ~indinqReqion ARCRGFLMRV AQCRGYLARK AMCRGYLARK AVCRGYLARK LC20 BindinqRec~ion Ch Sk Ch Sm Rb Sm Ch NMB KLAEIITRTQ KITDVIIAFQ KITDVIMAFQ KITDIIIFFQ EYRRMVERRE SIFCIQYNVR SFMNVKHWPW AFAKRQQQLT A M K V I Q R N C A A Y L K L R N W Q W AFAKRQQQLT A M K V I Q R N C A A Y L K L R N W Q W AFAKKQQQLS A L K I L Q R N C A A Y L K L R H W Q W 831 839 833 833 Ch Sk Ch Sm Rb Sm Ch NMB MKLFFKIKPL WRLFTKVKPL WRLFTKVKPL WRVFTKVKPL ANMKEEFEKT KEELAKSEAK QAKDEELQRT KERQQKAEAE QAKEDELQKI KERQQKAESE QAKDEELMKVKEKQTKVEAE RKELEEKMVV LKELEQKHTQ LQELQQKHTQ LEEMERKHQQ 881 889 883 883 Ch Sk Ch Sm Rb Sm Ch NMB LLQEKNDLQL QVQAEADSLA DAEERCDQLI LCEEKNLLQE KLQAETELYA E A E E M R V R L A LSEEKNLLQE QLQAETELYA E A E E M R V R L A LLEEKNILAE QLQAETELFA E A E E M R A R L A KTKIQLEAKI AKKQELEEIL AKKQELEEIL AKKQELEEIL KEVTERAEDE HEMEARIEEE HEMEARLEEE HDLESRVEEE 931 939 933 933 Ch Sk Ch Sm Rb Sm Ch NMB EEINAELTAK KRKLEDECSE EERSQQLQAE KKKMQQQMLD EDRGQQLQAE RKKMAQQMLD EERNQILQNE KKKMQGHIQD LKKDIDDLEL LEEQLEEEEA LEEQLEEEEA LEEQLDEEEG TLAKVEKEKH A T E N K V K N L T A R Q K L Q L E K V TADGKIKKME A R Q K L Q L E K V TAEAKIKKLE A R Q K L Q L E K V STEAKIKKME 981 989 933 933 Ch Sk Ch Sm Rb Sm Ch NMB EEMAVLDETI AKLTKEKKAL DDILIMEDQN NKLTKERKLL DDILVMDDQN NKLSKERKLL EEILLLEDQN SKFLKEKKLM QEAHQQTLDD EERVSDLTTN EERISDLTTN EDRIAECTSQ LQVEEDKVNT LAEEEEKAKN LAEEEEKAKN LAEEEEKAKN LTKAKTKLEQ LTKLKNKHES LTKLKNKHES LAKLKNKQEM 1031 1039 1033 1033 Ch Sk Ch Sm Rb Sm Ch NMB QVDDLEGSLE QEKKLRMDLE MISELEVRLK KEEKSRQELE MISELEVRLK KEEKSRQELE MITDLEERLK KEEKTRQELE RAKRKLEGDL KLAHDSIMDL E N D K Q Q L D E K KIKRKLEGES SDLHEQIAEL Q A Q I A E L K A Q KLKRKMDGEA SDLHEQIADL QAQIAELKMQ KAKRKLDGET TDLQDQIAEL QAQIEELKIQ 1081 1089 1083 1083 Ch Sk Ch Sm Rb Sm Ch NMB LKKKDFEISQ IQSKIEDEQA LAKKEEELQA ALARLEDETS LAKKEEELQA ALARLEDETS LAKKEEELQA A L A R G D E E A V LGMQLQKKIK QKNNALKKIR QKNNALKKIR QKNNALKVIR ELQARIEELE E E I E A E R T S R ELESHISDLQ E D L E S E K A A R ELEGHISDLQ E D L D S E R A A R ELQAQIAELQ E D L E S E K A S R 1131 1139 1133 1133 Ch Sk Ch Sm Rb Sm Ch NMB AKAEKHRADL NKAEKQKRDL NKAEKQKRDL NKAEKQKRDL SRELEEISER SEELEALKTE GEELEALKTE SEELEALKTE LEEAGGATAA LEDTLDTTAT LEDTLDTTAT LEDTLDTTAA QIEMNKKREA QQELRAKREQ QQELRAKREQ QQELRTKREQ EFQKMRRDLE EVTVLKRALE EVTVLKKALD EVAELKKAIE 1181 1189 I183 i183 Ch Sk Ch Sm Rb Sm Ch NMB EATLQHEATA EETRTHEAQV EETRSHEAQV EETKNHEAQI AALRKKHADS QEMRQKHTQA QEMRQKHTQV QEIRQRHATA TAELGEQIDN LQRVKQKLEK EKSELKMEID VEELTEQLEQ FKRAKANLDK T K Q T L E K D N A VEELTEQLEQ FKRAKANLDK T K Q T L E K E N A LEELSEQLEQ AKRFKANLEK N K Q G L E S D N K 1231 1239 1233 1233 S-2 ~ g i o n ~ LKSAESEKEM LQVTRQEEEM LQVTRQEEEM LQVTRQEEEL Typical 28 residue repeat FIGURE 3 smooth and skeletal muscle myosin than is the site at loop 2 that has already been discussed.