Tallon M A, Shenbagamurthi P, Marcus S, Becker J M, Naider F
Department of Chemistry, The College of Staten Island, City University of New York 10301.
Biochemistry. 1987 Dec 1;26(24):7767-74. doi: 10.1021/bi00398a035.
The synthesis and biological activity are reported for extended analogues of the secreted tridecapeptide alpha-factor (Trp-His-Trp-Leu-Gln-Leu-Lys-Pro-Gly-Gln-Pro-Met-Tyr) from Saccharomyces cerevisiae. Peptides with Ala, Glu-Ala, Ala-Glu-Ala, or Glu-Ala-Glu-Ala attached to the amino terminus of alpha-factor were synthesized by the solid-phase method on a (phenylacetamido)methyl (PAM) resin, using a combination of dicyclohexylcarbodiimide- and 1-hydroxybenzotriazole-accelerated active ester coupling procedures. Free peptides were obtained by hydrogen fluoride (HF) cleavage in the presence of appropriate scavengers. Normal high HF cleavage and "low-high" HF cleavage were equally effective in liberating the desired product from the PAM resin. Yields of pure peptide ranged from 9% to 17%. All of the extended alpha-factors, which represent sequences of pro-alpha-factor coded for in the MF alpha 1 structural gene, caused morphological aberrations (shmoo assay) in strain X2180-1A (MATa) the same as those caused by the tridecapeptide. The 14-peptide was equally active compared to the native alpha-factor whereas the 17-peptide was 5-10-fold less active. The analogues also arrested to various degrees (halo assay) the growth of S. cerevisiae RC629 (MATa sst1) and S. cerevisiae RC631 (MATa sst2), two supersensitive mutants, and were converted to pheromones of equal activity by treatment with V8 protease. A temperature-sensitive receptor mutant responded to all the peptides at the permissive but not the restrictive temperature. An alpha-factor antagonist, des-Trp1,Ala3-alpha-factor, inhibited activity of all extended peptides.(ABSTRACT TRUNCATED AT 250 WORDS)
本文报道了酿酒酵母分泌型十三肽α-因子（色氨酸-组氨酸-色氨酸-亮氨酸-谷氨酰胺-亮氨酸-赖氨酸-脯氨酸-甘氨酸-谷氨酰胺-脯氨酸-甲硫氨酸-酪氨酸）扩展类似物的合成及其生物活性。在（苯乙酰氨基）甲基（PAM）树脂上，采用二环己基碳二亚胺和1-羟基苯并三唑加速的活性酯偶联程序相结合的固相法，合成了在α-因子氨基末端连接丙氨酸、谷氨酸-丙氨酸、丙氨酸-谷氨酸-丙氨酸或谷氨酸-丙氨酸-谷氨酸-丙氨酸的肽。在适当的清除剂存在下，通过氟化氢（HF）裂解获得游离肽。正常的高HF裂解和“低-高”HF裂解在从PAM树脂上释放所需产物方面同样有效。纯肽的产率在9%至17%之间。所有扩展的α-因子，代表MFα1结构基因编码的前α-因子序列，在X2180-1A（MATa）菌株中引起形态畸变（shmoo试验），与十三肽引起的相同。与天然α-因子相比，14肽的活性相同，而17肽的活性低5至10倍。这些类似物还不同程度地（晕圈试验）抑制了酿酒酵母RC629（MATa sst1）和酿酒酵母RC631（MATa sst2）这两个超敏感突变体的生长，并通过V8蛋白酶处理转化为活性相等的信息素。一个温度敏感的受体突变体在允许温度下对所有肽有反应，但在限制温度下无反应。一种α-因子拮抗剂，去色氨酸1，丙氨酸3-α-因子，抑制所有扩展肽的活性。（摘要截短于250字）
