POLYGLUTAMYLATED ALPHA-TUBULIN CAN ENTER THE TYROSINATION DETYROSINATION CYCLE

被引:42
作者
EDDE, B
ROSSIER, J
LECAER, JP
PROME, JC
DESBRUYERES, E
GROS, F
DENOULET, P
机构
[1] CNRS, CTR RECH BIOCHIM & GENET CELLULAIRE, F-31062 TOULOUSE, FRANCE
[2] CNRS, PHYSIOL NERVEUSE LAB, F-91198 GIF SUR YVETTE, FRANCE
关键词
D O I
10.1021/bi00117a014
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have previously identified a major modification of neuronal alpha-tubulin which consists of the posttranslational addition of a varying number of glutamyl units on the gamma-carboxyl group of glutamate residue 445. This modification, called polyglutamylation, was initially found associated with detyrosinated alpha-tubulin [Edde, B., Rossier, J., Le Caer, J. P., Desbruyeres, E., Gros, F., & Denoulet, P. (1990) Science 247, 83-85]. In this report we show that a lateral chain of glutamyl units can also be present on tyrosinated alpha-tubulin. Incubation of cultured mouse brain neurons with radioactive tyrosine, in the presence of cycloheximide, resulted in a posttranslational labeling of six alpha-tubulin isoelectric variants. Because both tyrosination and polyglutamylation occur in the C-terminal region of alpha-tubulin, the structure of this region was investigated. [H-3]tyrosinated tubulin was mixed with a large excess of unlabeled mouse brain tubulin and digested with thermolysin. Five peptides, detected by their radioactivity, were purified by high-performance liquid chromatography. Amino acid sequencing and mass spectrometry showed that one of these peptides corresponds to the native C-terminal part of alpha-tubulin (VEGEGEEEGEEY)-V-440-Y-451 and that the remainders bear a varying number of glutamyl units linked to glutamate residue 445, which explains the observed heterogeneity of tyrosinated alpha-tubulin. A quantitative analysis showed that the different tyrosinated forms of alpha-tubulin represent a minor (13%) fraction of the total alpha-tubulin present in the brain and that most (80%) of these tyrosinated forms are polyglutamylated. The different forms of alpha-tubulin were found to be equal substrates for tubulin tyrosine ligase and tubulin carboxypeptidase, which indicates that alpha-tubulin can enter the tyrosination/detyrosination cycle independently of its degree of glutamylation.
引用
收藏
页码:403 / 410
页数:8
相关论文
共 43 条
[1]   CHARACTERIZATION OF POSTTRANSLATIONAL MODIFICATIONS IN NEURON-SPECIFIC CLASS-III BETA-TUBULIN BY MASS-SPECTROMETRY [J].
ALEXANDER, JE ;
HUNT, DF ;
LEE, MK ;
SHABANOWITZ, J ;
MICHEL, H ;
BERLIN, SC ;
MACDONALD, TL ;
SUNDBERG, RJ ;
REBHUN, LI ;
FRANKFURTER, A .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (11) :4685-4689
[2]   INDIVIDUAL MICROTUBULES IN THE AXON CONSIST OF DOMAINS THAT DIFFER IN BOTH COMPOSITION AND STABILITY [J].
BAAS, PW ;
BLACK, MM .
JOURNAL OF CELL BIOLOGY, 1990, 111 (02) :495-509
[3]   SOME COMMON PROPERTIES OF PROTEIN THAT INCORPORATES TYROSINE AS A SINGLE UNIT AND MICROTUBULE PROTEINS [J].
BARRA, HS ;
ARCE, CA ;
RODRIGUEZ, JA ;
CAPUTTO, R .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1974, 60 (04) :1384-1390
[4]   POSTTRANSLATIONAL TYROSINATION DETYROSINATION OF TUBULIN [J].
BARRA, HS ;
ARCE, CA ;
ARGARANA, CE .
MOLECULAR NEUROBIOLOGY, 1988, 2 (02) :133-153
[5]   TOTAL TUBULIN AND ITS AMINOACYLATED AND NON-AMINOACYLATED FORMS DURING THE DEVELOPMENT OF RAT-BRAIN [J].
BARRA, HS ;
ARCE, CA ;
CAPUTTO, R .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1980, 109 (02) :439-446
[6]   NA+-CHANNEL-ASSOCIATED SCORPION TOXIN RECEPTOR-SITES AS PROBES FOR NEURONAL EVOLUTION INVIVO AND INVITRO [J].
BERWALDNETTER, Y ;
MARTINMOUTOT, N ;
KOULAKOFF, A ;
COURAUD, F .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1981, 78 (02) :1245-1249
[7]  
DAHL JL, 1979, BIOCHEM BIOPH RES CO, V86, P822, DOI 10.1016/0006-291X(79)91786-8
[8]   EVOLUTION OF TUBULIN HETEROGENEITY DURING MOUSE-BRAIN DEVELOPMENT [J].
DENOULET, P ;
EDDE, B ;
JEANTET, C ;
GROS, F .
BIOCHIMIE, 1982, 64 (03) :165-172
[9]   DIFFERENTIAL EXPRESSION OF SEVERAL NEUROSPECIFIC BETA-TUBULIN MESSENGER-RNAS IN THE MOUSE-BRAIN DURING DEVELOPMENT [J].
DENOULET, P ;
EDDE, B ;
GROS, F .
GENE, 1986, 50 (1-3) :289-297
[10]  
DENOULET P, 1988, STRUCTURE FUNCTIONS, P231