A strong protein‐tyrosine kinase activity is associated with a baculovirus‐expressed chicken <i>tkl</i> gene

Gärtner, Thomas; Raab, Gerhard; Raab, Monika; Strebhardt, Klaus; Rübsamen‐Waigmann, Helga

doi:10.1111/j.1432-1033.1992.tb17162.x

Cited by 6 publications

(3 citation statements)

References 47 publications

(29 reference statements)

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“…Reiss et al (23) showed that GAPDH served as substrate for purified epidermal growth factor receptor kinase. It was also demonstrated by phosphoamino acid analysis that GAPDH was tyrosine phosphorylated by tyrosine kinase related to lymphode cell kinase (29). Likewise, Sergienko et al (24) found that rabbit muscle GAPDH contained phosphotyrosine.…”

Section: Discussionmentioning

confidence: 92%

A GAPDH Mutant Defective in Src‐Dependent Tyrosine Phosphorylation Impedes Rab2‐Mediated Events

Tisdale

Artalejo

2007

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Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has multiple intracellular activities in addition to its role in gluconeogenesis. Indeed, we have reported that GAPDH is required for Rab2-mediated retrograde transport from vesicular tubular clusters (VTCs). These diverse GAPDH activities are the result of posttranslational modifications that confer a new function to the enzyme. In that regard, GAPDH is tyrosine phosphorylated by Src. To establish the functional significance of this modification for GAPDH activity in Rab2-dependent events, an amino acid substitution was made at tyrosine 41 (GAPDH Y41F). The inability of Src to phosphorylate purified recombinant GAPDH Y41F was confirmed in an in vitro kinase assay. The mutant was then employed in a quantitative membrane-binding assay that measures Rab2 recruitment of soluble components to VTCs. As we observed with GAPDH wild type, Rab2 promoted GAPDH Y41F binding to membranes in a dose-dependent manner, indicating that GAPDH tyrosine phosphorylation is not required for VTC association. However, GAPDH was tyrosine phosphorylated on VTCs. Importantly, GAPDH Y41F blocked vesicular stomatitis virus-G transport in an assay that reconstitutes endoplasmic reticulum to Golgi trafficking, indicating that phosphorylation of tyrosine 41 is essential for GAPDH activity in the early secretory pathway. The block in transport is because of the decreased binding of atypical protein kinase C i/l to GAPDH Y41F, which reduces b-coat protein association with the VTC and subsequent formation of Rab2-mediated retrograde vesicles. Our results suggest that Src plays a pivotal role in regulating the interaction of Rab2 effectors on the VTC.

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Section: Discussionmentioning

confidence: 92%

A GAPDH Mutant Defective in Src‐Dependent Tyrosine Phosphorylation Impedes Rab2‐Mediated Events

Tisdale

Artalejo

2007

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“…Во-первых, модификация поверхностных остатков белковой молекулы, которая влияет на взаимодействие ГАФД с белкамипартнерами. Прежде всего, к этому типу относится фосфорилирование сериновых и тирозиновых остатков, приводящее к изменению связывания ГАФД с другими белками [12][13][14], а также гликирование лизиновых остатков, вызывающее инактивацию фермента и изменение некоторых его свойств [15,16]. Такой тип модификаций имеет важное значение для осуществления негликолитических функций ГАФД, из-за которых этот фермент получил поэтичное название «Moonlighting protein», не очень красиво переведенное на русский язык как «белок подработки» [17].…”

Section: пост-трансляционные модификации гафдunclassified

Posttranslational Modifications of the Sulfhydryl Group of the Cysteine Residue of Glyceraldehyde-3-Phosphate Dehydrogenase

Muronetz,

Medvedeva,

Schmalhausen

2024

Lomonosov Chemistry Journal

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This review considers the main types of oxidative posttranslational modi cations of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) targeting the sulfhydryl group of the catalytic cysteine residue Cys152. The highly reactive sulfhydryl group of Cys152 in the active centre of GAPDH undergoes oxidation and S-nitrosylation, leading to enzyme inactivation and destabilization. Upon reversible oxidation of the sulfhydryl group to form cysteine-sulfenic acid, the enzyme loses dehydrogenase activity, but gains the ability to catalyze the acyl-phosphatase reaction. Hydrolysis of the product of the dehydrogenase reaction, 1,3-diphosphoglycerate, under the action of the oxidized GAPDH leads to uncoupling of oxidation and phosphorylation at this stage of glycolysis. The action of nitric oxide results in S-nitrosylation of Cys152 GAPDH and the subsequent formation of cysteine-sulfenic acid due to hydrolysis of the S-NO-group. Data are presented on the relationship between S-nitrosylation of the catalytic Cys152 of GAPDH and its oxidation followed by S-glutathionylation of the enzyme at Cys152. The role of posttranslational modi cations of the sulfhydryl group of the catalytic cysteine residue in the regulation of enzyme activity, as well as the mechanisms ensuring the reversibility of such modi cations are discussed.

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“…The ®rst was its inability to discriminate between myristoylated, properly modi®ed recombinant c-Src proteins localized to infected insect cell membranes and unmodi®ed, denatured recombinant c-Src found in both soluble and insoluble states in these cells. Several groups have noted that 50% or more of baculovirusexpressed Src protein is not myristoylated (Gartner et al, 1992;Piwnica-Worms et al, 1990;Ramer et al, 1991). The second drawback is the requirement for using pH 11 bu er to elute c-Src from the column, which inevitably causes some denaturation.…”

Section: Expression and Puri®cation Of C-src From Insect Cellsmentioning

confidence: 99%

The PDGF receptor phosphorylates Tyr 138 in the c-Src SH3 domain in vivo reducing peptide ligand binding

Broome

Hunter

1997

Oncogene

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Treatment of quiescent NIH3T3 cells with PDGF BB results in the transient activation and hyperphosphorylation of the protein-tyrosine kinase, c-Src. These e ects correlate with novel serine and tyrosine phosphorylations in the N-terminal non-catalytic region of the molecule, which contains an SH3 and SH2 domain. In this study, a site of PDGF-induced tyrosine phosphorylation was mapped to Tyr 138 in the SH3 domain; Tyr 138 is exposed on the SH3 peptide binding surface. This same site is phosphorylated in vitro by the PDGF receptor when puri®ed baculovirus-expressed c-Src is complexed with the activated receptor. Phosphorylation of Tyr 138 required association of c-Src with the PDGF receptor via its SH2 domain. When a c-Src Phe 138 mutant was stably expressed in Src 7 mouse ®broblasts, it was activated to the same extent as wild type c-Src following PDGF stimulation, indicating that phosphorylation of this site is not required for PDGF-mediated activation. However, Tyr 138 phosphorylation was found to diminish SH3 domain peptide ligand binding ability in vitro.

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A strong protein‐tyrosine kinase activity is associated with a baculovirus‐expressed chicken tkl gene

Cited by 6 publications

References 47 publications

A GAPDH Mutant Defective in Src‐Dependent Tyrosine Phosphorylation Impedes Rab2‐Mediated Events

A GAPDH Mutant Defective in Src‐Dependent Tyrosine Phosphorylation Impedes Rab2‐Mediated Events

Posttranslational Modifications of the Sulfhydryl Group of the Cysteine Residue of Glyceraldehyde-3-Phosphate Dehydrogenase

The PDGF receptor phosphorylates Tyr 138 in the c-Src SH3 domain in vivo reducing peptide ligand binding

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