Interaction of C-terminal truncated human alphaA-crystallins with target proteins.
Interaction of C-terminal truncated human alphaA-crystallins with target proteins.
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C末端截短的人α-晶状体与靶蛋白的相互作用。
DOI：
10.1371/journal.pone.0003175
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发表时间：
2008-09-09
期刊：
影响因子：
3.7
通讯作者：
Abraham, Edathara C.
中科院分区：
文献类型：
--
作者：
Kumarasamy, Anbarasu;Abraham, Edathara C.
在人类晶状体中，αA-晶体蛋白的很大一部分存在于第172、168和162位残基上的C末端残基上。以乙醇脱氢酶和βL晶体蛋白为靶蛋白测定，αA1-172中伴侣活性增加，αA1-168和αA1-162中伴侣活性降低。这项研究的目的是表明C末端残基的缺失是否会影响蛋白质与靶蛋白质的相互作用。我们的假设是，伴侣-靶蛋白结合动力学，也称为亚单位交换率，预计将反映伴侣活性的变化。为了研究这一点，我们依赖于利用胺和半胱氨酸特异的荧光探针的荧光共振能量转移(FRET)。Adh和αA1-172的亚基交换率(K)与adh和αA-wt相近，αA1-168较低，αA1-162的k值最低。以βL晶体蛋白为靶蛋白时，αA1-172的k值略高于αA-wt，αA1-168和αA1-162的k值较低。正如先前研究所预期的那样，αA1-172的伴侣活性略好于αA-wt，αA1-168的伴侣活性与αA-wt相似，而αA1-162的伴侣活性显著降低。包括Arg-163和C末端柔性臂在内的11个C末端残基的切割显著影响了与靶蛋白的相互作用。以亲水性为主的柔性臂似乎是保持伴侣-靶蛋白复合体溶解所必需的。
Significant portion of αA-crystallin in human lenses exists as C-terminal residues cleaved at residues 172, 168, and 162. Chaperone activity, determined with alcohol dehydrogenase (ADH) and βL-crystallin as target proteins, was increased in αA1–172 and decreased in αA1–168 and αA1–162. The purpose of this study was to show whether the absence of the C-terminal residues influences protein-protein interactions with target proteins. Our hypothesis is that the chaperone-target protein binding kinetics, otherwise termed subunit exchange rates, are expected to reflect the changes in chaperone activity. To study this, we have relied on fluorescence resonance energy transfer (FRET) utilizing amine specific and cysteine specific fluorescent probes. The subunit exchange rate (k) for ADH and αA1–172 was nearly the same as that of ADH and αA-wt, αA1–168 had lower and αA1–162 had the lowest k values. When βL-crystallin was used as the target protein, αA1–172 had slightly higher k value than αA-wt and αA1–168 and αA1–162 had lower k values. As expected from earlier studies, the chaperone activity of αA1–172 was slightly better than that of αA-wt, the chaperone activity of αA1–168 was similar to that of αA-wt and αA1–162 had substantially decreased chaperone activity. Cleavage of eleven C-terminal residues including Arg-163 and the C-terminal flexible arm significantly affects the interaction with target proteins. The predominantly hydrophilic flexible arm appears to be needed to keep the chaperone-target protein complex soluble.
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影响因子：
2.9
作者：
Bera, S;Abraham, EC
通讯作者：
Abraham, EC
影响因子：
3.4
作者：
Ma, ZX;Hanson, SRA;Smith, JB
通讯作者：
Smith, JB
影响因子：
2
作者：
TAKEMOTO, LJ
通讯作者：
TAKEMOTO, LJ
影响因子：
2.9
作者：
Aziz, Atya;Santhoshkumar, Puttur;Abraham, Edathara C.
通讯作者：
Abraham, Edathara C.
DOI：
10.1073/pnas.79.7.2360
发表时间：
1982-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
INGOLIA, TD;CRAIG, EA
通讯作者：
CRAIG, EA