TY - JOUR
T1 - Complete primary structure of vertebrate smooth muscle myosin heavy chain deduced from its complementary DNA sequence. Implications on topography and function of myosin
AU - Yanagisawa, Masashi
AU - Hamada, Yoshio
AU - Katsuragawa, Yoshinari
AU - Imamura, Michihiro
AU - Mikawa, Takashi
AU - Masaki, Tomoh
N1 - Funding Information:
Tn our model. both the ATT’-binding site and the atc~tin~hindiny sit’e are presumably composed of more than one domain: the ATP sit,e is situated t)et)ween the 2.iK and the 5OK subfragment domains. while t Irt~6 OK and thcb2 OR suttfragrntbnts c7)rtstitut.e ttifl actin site. Further. thts individual domains are thought t.o be rssentiall>~ rigid but, linked t.ogethrr 1)~ flrxihle pqrtit-l~~ segments (Bennett & Hubcr. 1984). ATT’-induced changes in the shape of $1 have ht~i ohs;rrvCld t)!* electron micaroscaopy of ~~hemioally crosslinked a,c+in--Sl cbompleses (Craig rt crl.. 1985). Transient electrical birefringence tneasttrements on Sl have indicat’ed that SI has at least two flexibly at’tached tertiary structural domains (Highsmith & Eden, 1986). Further. thri binding of nu&ot~ide and/or a&in to Kl :iffecbts thtb suscsrptihility of thr 25Kj5OK and ;sOKMK ,jt,tn~tional peptides to protea,ses (Albplegate K-Keixler, 1983, 1984). Thaw olrxwvations, when taken in cwnjunction with the crontigurat ion of our model. suggest that t tit>d ianges in tjtie st& of the nucleotidc~ site (i.e. occupied by ATT’. ADP-P, or ADP. etc.) Would iriclucbta a rrsr,r;-tngclrnt~rlt of the two parallel r/b domains t~omposed of ttle 25K and 5OK subfrap-tnertts. much like the “.induced fit’” proposed in ftc>?cokinasr. f)I-tosphogl~(“ratr~ kinasrl and phos-phofructokinaw (Kennet t & Huber? 1984). The domain movement woultl IF transtnittcbd to other rvgiorts of Sl ittc+tding the a,ctin site. through H rnoditic*ntion of the relative positions of’ the t.hrey domains: and /~irp wrsu. the binding of’ aclin ma? tnodttlat.e the c.ottformatiorr of the .4TP site in a similar matitt~~r~ \Vr thiink Ihs Tatsuo Otri a,nd Rogw 31. Haigh for thv viwvful wading trt’ thtb manuwript: I)r Grnji hlatsuda for informxtion on the pptidt~ seyuenw of rhicaken sk&tal AIH ( q I,PfiW J)\~hlic%tion; and Miss H. Kajiura tirr twhnic~al a,ssistatrcbr in Jwpt~idv srquwxing. 1Z’e thank our ma.n~~ c~~llraptrs for their htimulating discussions of thcb topogtaph,I Of’ Sl. prticularly I)tY Ta kp-uki \Vakabltvashi. Fumi Morita. Kazuo Sutoh. \‘oh Okantot;,. Ktliich \yat~~,zmoto. Hirofumi Onishi and Takao Kodama. This work u-as sltpported 1)~ grant,s from thr Ministry of F:duc;~tiotr. Science and (‘ulture: the Ministy,. of’ HtAth and \V~lf:~w: Asahi-Shinbun ( IO.; I’rhatx ;Ilenitrrial I~oundation: and R~ewarch Foundation , It’ l_‘tliv,~rsit>~ of Tsrtkul)a. Following the c~otrtpletion of this v~~rk. NY Iri~rttrcl that thr pptidc! srquenc~ of the SK suhfragmwt of’ chic*ketl gizzard Sl had twvrl detrrminetl (Onishi r! ~1.. IWtI). \\‘r hitvv cwdirmecl that thr srque~~w is itlctltic~al \vith amino acitl rexidws fiifi to Xi8 in the r~I)S.\-t~tlc~c,clrfl ~;(‘(jllt’tlw’.
PY - 1987/11/20
Y1 - 1987/11/20
N2 - The 1979 amino acid sequence of embryonic chicken gizzard smooth muscle myosin heavy chain (MHC) hase been determined by cloning and sequencing its cDNA. Genomic Southern analysis and Northern analysis with the cDNA sequence show that gizzard MHC is encoded by a single-copy gene, and this gene is expressed in the gizzard and aorta. The encoded protein has a calculated Mr of 229 × 103, and can be divided into a long α-helical rod and a globular head. Only 32 to 33% of the amino acid residues in the rod and 48 to 49% in the head are conserved when compared with nematode or vertebrate sarcomeric MHC sequences. However, the seven residue hydrophobic periodicity, together with the 28 and 196 residue repeat of charge distribution previously described in nematode myosin rod, are all present in the gizzard myosin rod. Two of the trypsin-sensitive sites in gizzard light meromyosin have been mapped by partial peptide sequencing to 99 nm and 60 nm from the tip of the myosin tail, where these sites coincide with the two "hinges" for the 6S/10S transition. In the head sequence, several polypeptide segments, including the regions around the putative ATP-binding site and the reactive thiol groups, are highly conserved. These areas presumably reflect conserved structural elements important for the function of myosin. A multi-domain folding model of myosin head is proposed on the basis of the conserved sequences, information on the topography of myosin in the literature, and the predicted secondary structures. In this model, Mg2+ ATP is bound to a pocket between two opposing α β domains, while actin undergoes electrostatic interactions with lysine-rich surface loops on two other domains. The actin-myosin interactions are thought to be modulated through relative movements of the domains induced by the binding of ATP.
AB - The 1979 amino acid sequence of embryonic chicken gizzard smooth muscle myosin heavy chain (MHC) hase been determined by cloning and sequencing its cDNA. Genomic Southern analysis and Northern analysis with the cDNA sequence show that gizzard MHC is encoded by a single-copy gene, and this gene is expressed in the gizzard and aorta. The encoded protein has a calculated Mr of 229 × 103, and can be divided into a long α-helical rod and a globular head. Only 32 to 33% of the amino acid residues in the rod and 48 to 49% in the head are conserved when compared with nematode or vertebrate sarcomeric MHC sequences. However, the seven residue hydrophobic periodicity, together with the 28 and 196 residue repeat of charge distribution previously described in nematode myosin rod, are all present in the gizzard myosin rod. Two of the trypsin-sensitive sites in gizzard light meromyosin have been mapped by partial peptide sequencing to 99 nm and 60 nm from the tip of the myosin tail, where these sites coincide with the two "hinges" for the 6S/10S transition. In the head sequence, several polypeptide segments, including the regions around the putative ATP-binding site and the reactive thiol groups, are highly conserved. These areas presumably reflect conserved structural elements important for the function of myosin. A multi-domain folding model of myosin head is proposed on the basis of the conserved sequences, information on the topography of myosin in the literature, and the predicted secondary structures. In this model, Mg2+ ATP is bound to a pocket between two opposing α β domains, while actin undergoes electrostatic interactions with lysine-rich surface loops on two other domains. The actin-myosin interactions are thought to be modulated through relative movements of the domains induced by the binding of ATP.
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U2 - 10.1016/0022-2836(87)90302-0
DO - 10.1016/0022-2836(87)90302-0
M3 - Article
C2 - 2892941
AN - SCOPUS:0023661175
SN - 0022-2836
VL - 198
SP - 143
EP - 157
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 2
ER -