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氮代谢调控相关蛋白质泛素化修饰筛选
吕永坤,
周景文,
刘龙
k
(1
1
一
i
郭大学
IJ
二物[{'~"j!:院,江苏
)c
锡
214122)
摘要:通过对氮代谢相关透性酶及调控因子泛素化修饰的筛选,为通过改造蛋白质泛素化修饰
来解除氮代谢阻遏效应提供参考。对此前建立的一种蛋白质泛素化快速检测方法进行了改造和
发展,进一步增强了阳性子的荧光信号,并使得该方法可以用于定量分析,从而使泛素化检测的
结果更为可靠。对氮代谢物阻遏效应密切相关的
10
个蛋白质的检测结果表明,其中
5
个受到泛
素化修饰,
4
个不能受到泛素化修饰,
1
个不能确定是否受到泛素化修饰。除建立了一种可靠的
蛋白质泛素化检测方法,也为通过蛋白质泛素化调控途径解除氮代谢物阻遏效应提供了思路。
关键词:酿酒酵母;透性酶;调控因子;泛素化;检测
中图分类号
:S
665.2
文献标志码
:A
文章编号
:1673--1689(2014)06--0576--07
Ubiquitination Detection of NCR Related Permeases and Regulators
LV
Yongkun ,
ZHOU
Jingwen ,
LIU
Long
大
(School
o[
Biotechnology
,Jiangnan
University
,
Wuxi
214122 , China)
Abstract:
To
give
SUpp
Ol
t to
release
the
nitrogen catabolite repression
(NCR)
,by rebuiling the
ubiquitination system
,
the
ubiquitination
of NCR
related
permeases
and
regulators was detected. A
fast ubiquitination detection
procedure
established
before was modified
and
developed, which
enhanced
the
fluorescence of
the
positive control
and
could
quantify the fluorescence signal. The
fluorescence quantification made this
procedure
more reliable.
It
was
demonstrated
that
5 of the
NCR
related
proteins were
ubiquitinated
, 4 were not
ubiquitinated
and
1 was not determined. We
not only
established
a
reliable
ubiquitination
detection
procedure
,
but
also gave a
idea
to
release
NCR through
the
ubiquitination
modification
procedure.
Keywords:
5α
ccharomyces
cerevisiae ,
permease
, regulator ,
ubiquitination
,
detection
基于氮源的氮代谢调控影响到酿酒酵母
Sα
cch
α
romyces
cerevisiae (5.
cerevisiae)
几乎
10%
的
基因表达
Pl
。当培养基中同时存在多种氮源时,
S.
cerem
Sl
ae
会优先利用丰富型氮游、如谷氨酷胶、镀
收稿日期:
2013-10-09
盐、谷氨酸、天冬氨酷等,只有当培养基中不存在丰
富型氮塘、或者丰富型氮源被耗尽时
,
5.
cerems~ae
才
会开始利用精氨酸、尿素、尿囊素、
GABA
等贫乏型
氮源
O
这种由丰富型氮源引起的对贫乏型氮源利用
基金项目.国家
973
计划项口
(2012CB720800)
;同家白然科学基金重点项口
(31130043)ν
*通信作者·文Ij
)主
(1980-)
,刃,江苏元锡人
J
学博|二.副教授.硕
1:
研究
'1
导师主安从事做
't
物系统代谢工程及工业酶的分
F
定
/r'J11
Í\ó'i
{ri
研究。
E-mail:
IO
l1
gliu@jia
l1
~a
l1
.edu.cn
576
Journal
of
Food
Science
and
Bi
otechnology
Vυ
1.3
3
' .6
2014