The covalent structure of pig kidney fructose 1,6-bisphosphatase: sequence of the 60-residue NH2-terminal peptide produced by digestion with subtilisin.
The covalent structure of pig kidney fructose 1,6-bisphosphatase: sequence of the 60-residue NH2-terminal peptide produced by digestion with subtilisin.
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猪肾果糖 1,6-双磷酸酶的共价结构：用枯草杆菌蛋白酶消化产生的 60 个残基 NH2 末端肽的序列。
DOI：
10.1016/0003-9861(81)90330-1
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发表时间：
1981
影响因子：
3.9
通讯作者：
Heinrikson,RL
中科院分区：
文献类型：
--
作者：
Marcus,F;Edelstein,I;Saidel,LJ;Keim,PS;Heinrikson,RL
用枯草杆菌蛋白酶消化天然猪肾果糖1，6-二磷酸酶四聚体，切割35，000分子量亚基中的每一个，产生两个主要片段：S-亚基（Mrca.29，000）和S-肽（Mr 6，500）。已确定S肽的以下氨基酸序列：AcThr → Asp → Gln → Ala → Phe → Asp → Thr → Asn → Ile →瓦尔→ Thr → Leu → Thr → Arg → Phe → Val → Met → Glu → Gln → Gly → Arg → Lys → Ala → Arg → Gly → Thr → Gly → Glu → Met → Thr → Gln → Leu → Leu → Asn → Ser → Leu → Cys → Thr → Ala → Val → Lys → Ser → Thr → Ala → Val → Lys → Ser → Thr → Ala z.sbnd;瓦尔→ Arg → Lys → Ala → Gly → Ile → Ala → His → Leu → Tyr → Gly → Ile → Ala。该序列与来自兔肝的酶的NH 2-末端60个残基的序列的比较（El-Dorryet al.，1977，Arch.Biochem.Biopolymer.182，763）揭示了与52个相同位置的强同源性和从残基26到60的序列的绝对同一性。尽管果糖1，6-二磷酸酶的枯草杆菌蛋白酶切割导致酶对AMP抑制的敏感性降低，但我们在测序的S-肽中没有发现AMP抑制相关的氨基酸残基。在吡哆醛-P修饰酶时发生的AMP敏感性的丧失不会导致S-肽中赖氨酰残基的修饰。无论是光亲和标记的果糖1，6-二磷酸酶与8-叠氮基-AMP，也不修改的半胱氨酰残基近端的AMP变构位点导致在位于NH 2-末端的60个氨基酸的肽的残基的修改。
Digestion of the native pig kidney fructose 1,6-bisphosphatase tetramer with subtilisin cleaves each of the 35,000-molecular-weight subunits to yield two major fragments: the S-subunit (Mrca.29,000), and the S-peptide (Mr6,500). The following amino acid sequence has been determined for the S peptide: AcThrAspGlnAlaAlaPheAspThrAsnIle Val ThrLeuThrArgPheValMetGluGlnGlyArgLysAla ArgGlyThrGlyGlu MetThrGlnLeuLeuAsnSerLeuCysThrAlaValLys AlaIleSerThrAla z.sbnd;ValArgLysAlaGlyIleAlaHisLeuTyrGlyIleAla. Comparison of this sequence with that of the NH2-terminal 60 residues of the enzyme from rabbit liver (El-Dorryet al., 1977,Arch. Biochem. Biophys.182, 763) reveals strong homology with 52 identical positions and absolute identity in sequence from residues 26 to 60.Although subtilisin cleavage of fructose 1,6-bisphosphatase results in diminished sensitivity of the enzyme to AMP inhibition, we have found no AMP inhibition-related amino acid residues in the sequenced S-peptide. The loss of AMP sensitivity that occurs upon pyridoxal-P modification of the enzyme does not result in the modification of lysyl residues in the S-peptide. Neither photoaffinity labeling of fructose 1,6-bisphosphatase with 8-azido-AMP nor modification of the cysteinyl residue proximal to the AMP allosteric site resulted in the modification of residues located in the NH2-terminal 60-amino acid peptide.