Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “phosphorylation”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 217 records · Page 12Linked to original sources
PubMed1977

The phosphorylation sites of troponin T from white skeletal muscle and the effects of interaction with troponin C on their phosphorylation by phosphorylase kinase.

1. The phosphorylation of troponin T from rabbit white sketetal muscle is catalysed by phosphorylase kinase, but not at a significant rate by bovine 3':5'-cyclic AMP-dependent protein kinase. 2. The amino acid sequences adjacent to the three major phosphorylation sites of troponin T were determined. 3. The serine in the N-terminal peptide (Asx,SerP, Glx)Glu-Val-Glu, is that phosphorylated (SerP, phosphoserine) when the troponin complex is isolated. 4. The other two sites of phosphorylation are located in the sequence Ala-Leu-(Ser, SerP)-Met-Gly-Ala-Asn-Tyr(Ser,SerP)Tyr. 5. When troponin T is phosphorylated in the presence of troponin C, the extent of phosphorylation at each site is considerably decreased. 6. CNBr fragments of troponin T are also phosphorylated by phosphorylase kinase, but the rate of phosphorylation at each site in the CNBr fragments is considerably slower than in the native protein. 7. From these studies it is suggested that troponin C interacts with troponin T in the region containing the two closely situated phosphorylation sites.

Amino Acid Sequence↗
PubMed1975

Phosphorylation of solubilized sarcoplasmic reticulum by orthophosphate and its thermodynamic characteristics. The dominant role of entropy in the phosphorylation.

A large fraction of the Ca-2plus- and Mg-2plus-dependent ATPase (EC 3.6.1.3) in sarcoplasmic reticulum membranes solubilized with Triton X-100 was phosphorylated with Pi. The phosphorylation required Mg-2plus but was strongly inhibited by low concentrations of Ca-2plus. A Ca-2plus ion concentration of 30 muM caused half-maximum inhibition in the presence of 50 mM MgCl2. The phosphorylated enzyme showed a rapid turnover and was in dynamic equilibrium with Pi in the medium. At equilibrium the amount of the phosphorylated enzyme increased markedly with increased in the reaction temperature. The apparent standard free energy change, the apparent standard enthalpy change, and the apparent standard entropy change in the formation of the phosphorylated enzyme from the enzyme-phosphate complex in the presence of excess Mg-2plus at 37 degrees and pH 7.0 were, respectively, 0.35 Cal per mol, 15.9 Cal per mol, and 50.2 e.u. per mol. The susceptibility of the acid-denatured phosphorylated enzyme to hydroxylamine showed that the phosphorylated enzyme is of an acyl phosphate type. The present results are consistent with the probability that the phosphorylation results from reversal of late steps in the Ca-2plus transport process. The results clearly show that the phosphorylated enzyme is stabilized by a great increase in entropy upon its formation from the enzyme-phosphate complex.

Adenosine Triphosphatases↗
PubMed2004

Septin 2 phosphorylation: theoretical and mass spectrometric evidence for the existence of a single phosphorylation site in vivo.

Septins constitute a family of conserved proteins that are required for cytokinesis in a wide range of organisms. Most cells express a set of septin proteins and these are found to assemble into hetero-oligomeric septin complexes that appear filamentous. However, the mechanisms controlling the function and polymerization of septins are not known. We therefore examined the possibility that septins could be post-translationally modified by phosphorylation. We present herein a combined theoretical and experimental approach for the analysis of Septin 2 (Sept2) monophosphorylation in vivo. We purified and characterized the human recombinant Sept2, a 45-kDa protein, expressed from Sf21 insect cells. Analysis by matrix-assisted laser desorption/ionization quadrupole time-of-flight mass spectrometry on the full-length protein sequence of wild-type Sept2 revealed a unique phosphorylation site at residue Ser248 in vivo, which is consistent with one of the twelve phosphorylation sites in the protein sequence theoretically predicted by the Netphos program. Additional predictions with the motif scan programs Scansite and Prosite suggest that the phosphorylation of wild-type Sept2 might be a potential substrate for casein kinase 2. Site-directed mutagenesis of residue 248 from serine to alanine abrogated this phosphorylation. The location of phosphorylation in Sept2 differs from the sites predicted for cGMP-dependent protein kinase (PKG) phosphorylation in Septin 3, raising the possibility that different septins may undergo distinct phosphorylation events that could control their functions in important cellular processes such as neurotransmission or cytokinesis.

Amino Acid Sequence↗
PubMed1995

Ah receptor phosphorylation: localization of phosphorylation sites to the C-terminal half of the protein.

The aryl hydrocarbon receptor (AhR) is a transcriptional enhancer activated by the binding of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related xenobiotics. Ligand binding initiates a series of poorly understood molecular events which confers recognition of cis-acting elements located in regulatory regions of particular structural genes, such as CYP1A1. Several studies have suggested that AhR phosphorylation may be instrumental in activating the AhR to a DNA-binding state. In agreement with previous investigations, treatment of the AhR with acid phosphatase resulted in the loss of DNA-binding activity. To further evaluate the functional role of AhR phosphorylation we determined whether TCDD binding altered total AhR phosphorylation, and identified phosphorylated regions by the examination of chemical cleavage patterns. The AhR was isolated by immunoprecipitation from [32P]-orthophosphate-labeled Hepa 1 cells grown in the presence or absence of TCDD. Examination of the amount of 32P associated with the AhR indicated that the total level of AhR phosphorylation was not affected by ligand binding. Chemical cleavage with hydroxylamine and cyanogen bromide also revealed a similar pattern for liganded and unliganded AhR. The shortest regions of overlap determined by the chemical cleavage patterns localized phosphorylation sites to two regions in the C-terminal half of the AhR. One region is centrally located between amino acids 368 and 605 and within or adjacent to a DNA binding repressor domain. The other region is located at the glutamine-rich carboxyl terminus between amino acids 636 and 759. These data coupled with previous observations imply that total AhR phosphorylation is not altered by the ligand-elicited transformation to a DNA-binding form, but that phosphorylation nevertheless plays an important role in the ability of an active AhR-Arnt complex to associate with cis-acting regulatory elements.

Acid Phosphatase↗
PubMed1999

Tau phosphorylation by diisopropyl phosphorofluoridate (DFP)-treated hen brain supernatant inhibits its binding with microtubules: role of Ca2+/Calmodulin-dependent protein kinase II in tau phosphorylation.

Diisopropyl phosphorofluoridate (DFP) produces organophosphorus ester-induced delayed neurotoxicity (OPIDN) in hen, human, and other sensitive species. This is characterized by mild ataxia, which progresses to severe ataxia or paralysis in a few days. Ultrastructurally, OPIDN is associated with the degeneration of axons in central and peripheral nervous systems. Bacterially expressed longest human tau protein (htau40) phosphorylated by DFP-treated hen brain supernatant showed a decrease in microtubule binding in a shorter time than that phosphorylated by control hen brain supernatant. The decrease in htau40-microtubule binding observed on htau40 phosphorylation by the recombinant Ca2+/calmodulin (CaM)-dependent protein kinase II (CaM kinase II) alpha-subunit showed that CaM kinase II present in brain supernatant could participate in tau phosphorylation even in the absence of Ca2+/CaM and decrease tau-microtubule binding. In addition, use of htau40 mutants, htau40m1 (Ala416) and htau40m6 (Asp416), suggested that replacement of Ser416 by neutral or acidic amino acid produced some change in htau40 conformation that caused diminished binding with microtubules phosphorylated by brain supernatant in the presence of ethylene glycol bis(beta-aminoethyl ether) N, N'tetraacetic acid (EGTA). The change in conformation produced by Ser416 phosphorylation, however, was different from that produced by mutants since only nonmutated htau40 showed a significant decrease in binding with microtubules on phosphorylation by recombinant CaM kinase II in the presence of Ca2+/CaM compared to that obtained by phosphorylation in the presence of EGTA. This study showed that enhanced Ca2+/CaM-dependent protein kinase activity in DFP-treated hen brain supernatant may cause decreased tau-microtubule binding and destabilization of microtubules and may be involved in axonal degeneration in OPIDN.

Amino Acid Substitution↗
PubMed2002

Identification of a phosphorylation site on skeletal muscle myosin light chain kinase that becomes phosphorylated during muscle contraction.

A protein phosphorylated efficiently in vitro by MAP kinase-activated protein kinase-2 (MAPKAP-K2) was purified from skeletal muscle extracts and identified as the calcium/calmodulin-dependent myosin light chain kinase (MLCK). The phosphorylation site was mapped to Ser(161), a residue shown previously to be autophosphorylated by MLCK. The residue equivalent to Ser(161) became phosphorylated in vivo when rat hindlimbs were stimulated electrically. However, phosphorylation was triggered within seconds, whereas activation of MAPKAP-K2 required several minutes. Moreover, contraction-induced Ser(161) phosphorylation was similar in wild-type or MAPKAP-K2-/- mice. These results indicate that contraction-induced phosphorylation is probably catalyzed by MLCK and not MAPKAP-K2. Ser(161) phosphorylation induced the binding of MLCK to 14-3-3 proteins, but did not detectably affect the kinetic properties of MLCK. The sequence surrounding Ser(161) is unusual in that residue 158 is histidine. Previously, an arginine located three residues N-terminal to the site of phosphorylation was thought to be critical for the specificity of MAPKAP-K2.

14-3-3 Proteins↗
PubMed1999

DP1 phosphorylation in multimeric complexes: weaker interaction with cyclin A through the E2F1 cyclin A binding domain leads to more efficient phosphorylation than stronger interaction through the p107 cyclin A binding domain.

Stable enzyme-substrate interaction has been recognized as a major mechanism underlying the substrate preferences of cyclin-dependent kinases (Cdks). To learn the relationship between stability of physical association and efficiency of phosphorylation, we studied DP1 phosphorylation by cyclin A-Cdk2 in multiprotein complexes. When DP1 was connected to cyclin A-Cdk2 through E2F4 and p107, its phosphorylation was very inefficient, although its association with cyclin A-Cdk2 was stable. In contrast, DP1 was efficiently phosphorylated when weakly connected to cyclin A-Cdk2 via E2F1 or E2F4 with a fused cyclin A binding domain of E2F1. The transactivation activity of E2F4-DP1 heterodimers was reduced when DP1 was phosphorylated, while a phosphorylation deficient mutant of DP1 resisted this down-regulation. Phosphorylation and functional regulation of DP1 were not due to nuclear localization. Thus, stronger physical association between the kinase and the substrate does not necessarily lead to more efficient phosphorylation than weaker interaction does.

Amino Acid Sequence↗
PubMed2001

Y-27632, an inhibitor of Rho-associated kinases, prevents tyrosine phosphorylation of focal adhesion kinase and paxillin induced by bombesin: dissociation from tyrosine phosphorylation of p130(CAS).

A rapid increase in tyrosine phosphorylation of focal adhesion kinase (FAK), paxillin, and Crk-associated substrate (CAS) are prominent early events triggered by many G protein-coupled receptors (GPCRs), but the mechanisms involved remain unclear. Here, we examined whether the Rho-associated protein serine/threonine kinase family (ROCK) is a critical Rho effector in the pathway that links GPCR activation to the tyrosine phosphorylation of FAK, CAS, and paxillin. Treatment of Swiss 3T3 cells with Y-27632, a preferential inhibitor of ROCK, dramatically inhibited the formation of actin stress fibers, the assembly of focal contacts, and the increase in tyrosine phosphorylation of FAK and paxillin induced by bombesin in these cells. Surprisingly, we found that treatment with Y-27632 did not produce any detectable effect on bombesin-elicited CAS tyrosine phosphorylation even at the highest concentrations of Y-27632 tested. HA-1077, a preferential inhibitor of ROCK activity structurally unrelated to Y-27632, also attenuated the increase in the tyrosine phosphorylation of FAK and paxillin but did not affect the tyrosine phosphorylation of CAS induced by bombesin in Swiss 3T3 cells. The results demonstrate that ROCK-dependent tyrosine phosphorylation of FAK and paxillin can be dissociated from a ROCK-independent pathway leading to tyrosine phosphorylation of CAS.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1991

Qualitative and quantitative comparison of the distribution of phosphorylated and non-phosphorylated neurofilament epitopes within central and peripheral axons of adult hamster (Mesocricetus auratus).

The distribution of phosphorylated and non-phosphorylated neurofilament epitopes was determined immunocytochemically in adjacent 2 microns-thick sections of sciatic nerve, ventral root and spinal cord. Staining was scored as either intense, moderate or absent and the proportion of labeled axons was calculated for each category. Nearly all sciatic nerve and ventral root axons were immunoreactive with both antibodies against phosphorylated and non-phosphorylated neurofilaments and there were no significant differences in the number of intensely- or moderately-labeled axons. Within the spinal cord however, while the majority of large caliber axons was stained with both antibodies, there was a significant number of small caliber axons which stained only with antibodies against phosphorylated neurofilaments. These results show that phosphorylated and non-phosphorylated neurofilaments are extensively codistributed in CNS and PNS axons, and that in the CNS, staining intensity for non-phosphorylated epitopes is less in the smaller axons.

Animals↗
PubMed1991

Neurofilament immunoreactivity in populations of rat primary afferent neurons: a quantitative study of phosphorylated and non-phosphorylated subunits.

Neurofilament subunits in rat dorsal root ganglion (DRG) neurons were examined using five antibodies: NFH, RT97 and NFHP- recognise the 200 kDa subunit (NF200); NFH recognises both phosphorylated and non-phosphorylated forms of NF200 whereas RT97 and NFHP- are specific for the phosphorylated and non-phosphorylated forms respectively; 155 and anti-68 kD recognise the 155 kDa and 68 kDa subunits respectively. All the antibodies apart from NFHP- distinguished between the two populations of neurons corresponding to the light (L) and small dark (SD) cell types as previously shown for RT97. This demonstrates that L and SD neurons contain different levels of neurofilament and that the ability to discriminate between them is not unique to the antibody RT97. It is also evident that DRG neurons contain neurofilament composed of all three subunits. Since NFH and RT97, but not NFHP-, distinguished between the two populations, it appears that it is the presence of the phosphorylated form of NF200 that provides the basis for discrimination between the two cell types. After dephosphorylation of the neurofilament, NFHP- also discriminated between the two populations, indicating that there is more NF200 regardless of phosphorylation state in the L neurons. Observations made from unfixed DRGs indicate that all neurons contain some neurofilament and the neurofilament rich and neurofilament poor populations were also apparent. The use of colchicine apparently caused a small increase in neurofilament levels in at least some perikarya, presumably due to its blocking effect on axoplasmic transport. This caused some SD neurons to become neurofilament rich. We conclude that L neurons contain more neurofilament than SD neurons since both cell types contain non-phosphorylated NF200, but the L neurons also contain a much greater amount of the phosphorylated form.

Animals↗
PubMed1989

Multisite phosphorylation of microtubule-associated protein 2 (MAP-2) in rat brain: peptide mapping distinguishes between cyclic AMP-, calcium/calmodulin-, and calcium/phospholipid-regulated phosphorylation mechanisms.

Microtubule-associated protein 2 (MAP-2), a cytoskeletal protein of 280 kilodalton that is highly enriched in dendrites and neuronal perikarya, is subject to both cyclic AMP-, calcium/calmodulin-, and calcium/phospholipid-regulated phosphorylation when incubated with [gamma-32P]ATP in vitro. We have analyzed the different sites in MAP-2 phosphorylated by these three kinases in fresh or boiled cytosol from different regions of the rat brain, in particular the olfactory bulb, where only one form (MAP-2B) is present, and the cerebral cortex, where both forms (MAP-2A and MAP-2B) are equally enriched. Cyclic AMP-dependent protein kinase and calcium/calmodulin-dependent protein kinase II phosphorylated four common phosphorylation sites, as well as a number of distinct sites that were unique to each enzyme. Calcium/phospholipid-dependent protein kinase phosphorylated a minimum of 15 sites, only one of which appeared to be shared with the other protein kinases. Only serine residues were phosphorylated by cyclic AMP-dependent and calcium/phospholipid-dependent protein kinases, while both serine and threonine residues were phosphorylated by calcium/calmodulin-dependent protein kinase II. No differences were observed in the peptide maps of phospho-MAP-2 prepared from different brain regions. These results emphasize the complexity of the phosphorylation systems that may regulate the function of MAP-2 in situ.

Adenosine Triphosphate↗
PubMed1989

Purified maturation promoting factor phosphorylates pp60c-src at the sites phosphorylated during fibroblast mitosis.

We have previously shown that overexpressed chicken pp60c-src has retarded mobility, novel serine/threonine phosphorylation, and enhanced kinase activity during NIH 3T3 cell mitosis. Here we show that novel mitotic phosphorylations occur at Thr 34, Thr 46, and Ser 72. The possibility, previously raised, that Ser 17 is dephosphorylated during mitosis is excluded. The phosphorylated sites lie in consensus sequences for phosphorylation by p34cdc2, the catalytic component of maturation promoting factor (MPF). Furthermore, highly purified MPF from metaphase-arrested Xenopus eggs phosphorylated both wild-type and kinase-defective pp60c-src at these sites. Altered phosphorylation alone is sufficient to account for the large retardation in mitotic pp60c-src electrophoretic mobility: phosphorylation of normal pp60c-src by MPF retarded mobility and dephosphorylation of mitotic pp60c-src restored normal mobility. These results suggest that pp60c-src is one of the targets for MPF action, which may account in part for the pleiotropic changes in protein phosphorylation and cellular architecture that occur during mitosis.

Amino Acid Sequence↗
PubMed1994

The correlation between cortical neuron maturation and neurofilament phosphorylation: a developmental study of phosphorylated 200 kDa neurofilament protein in cat visual cortex.

Neurofilament proteins are major constituents of the neuronal cytoskeleton. When phosphorylated, neurofilament proteins are thought to be involved in slowing down the slower component of axonal transport, thereby increasing neurofilament stability and conferring resistance to proteolysis. In order to investigate the correlation between phosphorylated neurofilament and plasticity of the cat visual cortex, we applied a monoclonal antibody, which recognizes only the phosphorylated form of neurofilament protein, to kitten visual cortex. This antibody appeared to recognize only axons. There was a progressive increase of phosphorylated neurofilament-immunoreactive axons in the cortical layers and white matter during development. In the visual cortex of young kittens, phosphorylated neurofilament immunoreactive processes were mostly long, fine, and continuous axons. However, in kittens older than 2 months of age, phosphorylated neurofilament immunoreactivity was characterized by short, punctae-like staining. The immunoreactive axons were most dense in deep cortical layers V and VI initially, followed at later ages by layers IVa and IVb, and finally in layer IVc and in the superficial cortical layers of adult animals. This developmental laminar pattern of distribution was not affected by early input restriction and coincides with electrophysiological evidence of early neuronal maturation. In addition, the phosphorylated neurofilament immunoreactivity can also be found in the dendrites and perikaya of degenerating neurons. These results suggest that phosphorylated neurofilament can be used as an index of functional maturity in the kitten visual cortex and also as a marker for neurodegeneration.

Animals↗
PubMed1995

Analysis of phosphorylation of tau with antibodies specific for phosphorylation sites.

Previously, we determined sites of tau protein phosphorylation by tau protein kinase (TPK) I/glycogen synthase kinase 3 beta (GSK-3 beta) and TPKII/(cyclin-dependent kinase 5 (CDK5) + p23). We prepared antibodies specific for these sites of tau phosphorylated by TPKI and TPKII, using chemically synthesized phosphopeptides as antigens. Each antibody specifically reacts with each phosphorylation site. With these antibodies, it was confirmed that TPKI and TPKII are responsible for these phosphorylation sites, as reported previously, except that Ser404 is also weakly phosphorylated by TPKI alone. It was also observed that TPKII-phosphorylation enhances TPKI-phosphorylation. These results indicate that these antibodies are useful tools for investigation of the phosphorylation of tau by TPKI and TPKII.

Amino Acid Sequence↗
PubMed1991

Phosphorylation of basic fibroblast growth factor by purified protein kinase C and the identification of a cryptic site of phosphorylation.

We have further characterized the protein kinase C (PK-C) dependent phosphorylation of basic fibroblast growth factor (FGF). Intact recombinant basic FGF and a series of ten peptide fragments of basic FGF were phosphorylated by PK-C and the products were analyzed by SDS-PAGE and autoradiography. As expected, peptide fragments containing the known site of phosphorylation (Ser64) are substrates for phosphorylation. Surprisingly however, peptides containing the receptor binding domain of the mitogen [basic FGF(106-115)] are also phosphorylated. An examination of this sequence reveals the presence of a consensus sequence (Ser108-Ala109-Lys110) that mediates the reaction. Accordingly, all peptides that contain the core amino acids basic FGF(106-111) are substrates for phosphorylation. Peptide mapping of basic FGF confirms that Ser64 is the primary site of phosphorylation, suggesting that Ser108 is a cryptic consensus sequence. Because basic FGF is metabolized to sequence specific fragments after its binding and internalization into target cells, this cryptic site may in fact be phosphorylated in vivo.

Animals↗
PubMed2002

Protein kinase C phosphorylation of purified Na,K-ATPase: C-terminal phosphorylation sites at the alpha- and gamma-subunits close to the inner face of the plasma membrane.

The alpha-subunit of the Na,K-ATPase is phosphorylated at specific sites by protein kinases A and C. Phosphorylation by protein kinase C (PKC) is restricted to the N terminus and takes place to a low stoichiometry, except in rat. Here we show that the alpha-subunit of shark Na,K-ATPase can be phosphorylated by PKC at C-terminal sites to stoichiometric levels in the presence of detergents. Two novel phosphorylation sites are possible candidates for this PKC phosphorylation: Thr-938 in the M8/M9 loop located very close to the PKA site, and Ser-774, in the proximal part of the M5/M6 hairpin. Both sites are highly conserved in all known alpha-subunits, indicating a physiological role. A similar pattern of detergent-mediated phosphorylation by PKC was found in pig kidney Na,K-ATPase alpha-subunit. Interestingly, the kidney-specific gamma-subunit was phosphorylated by PKC in the presence of detergent. The close proximity of the novel PKC sites to the membrane suggests that targeting proteins to tether PKC into the membrane phase is important in controlling the in vivo phosphorylation of this novel class of membrane-adjacent PKC sites. It is suggested that in purified preparations where functional targeting may be impaired detergents are needed to expose the sites.

Amino Acid Sequence↗
PubMed1995

Phosphorylation of alpha-crystallin in rat lenses is stimulated by H2O2 but phosphorylation has no effect on chaperone activity.

Alpha crystallin (alpha), a phosphorylated structural protein of the lens, has been shown to be a chaperone preventing other lens proteins from aggregating. It is now demonstrated that with oxidative stress imposed on cultured rat lenses, the incorporation of labeled phosphate into the alpha polypeptide chains increased by two to four times over a 90-min period in comparison to control experiments. The phosphorylation rate of the B chain, alpha B, was twice that of the A chain, alpha A. However, phosphorylation of the alpha chains has an insignificant effect on the chaperone activity of alpha or the individual alpha A and alpha B chains as measured by suppressing the thermally induced aggregation of beta low or gamma crystallins. It was also found that the alpha A aggregates are more effective chaperones than the alpha B aggregates. The size of the macromolecules resulting from reaggregation of the isolated non-phosphorylated or phosphorylated alpha B chains are not markedly effected by phosphorylation. However, phosphorylation of the alpha A chain leads to a heterogeneous population with two major species, one similar in size to alpha A and another approximately twice as large. It is concluded that the phosphorylation of alpha is associated with some other function of the protein than that of chaperone activity and that this function may be linked to a protective response to oxidative stress.

Animals↗
PubMed1998

Effect of age on mitogen induced protein tyrosine phosphorylation in human T cell and its subsets: down-regulation of tyrosine phosphorylation of ZAP-70.

Several events of T cell activation have been reported to decline in humans with age. Since protein tyrosine phosphorylation is an early critical event of T cell activation, we performed a systematic analysis of the age-associated changes in the mitogen induced protein tyrosine phosphorylation of human T lymphocytes using SDS-PAGE and Western blotting techniques. Following stimulation with Con A and PHA, an identical pattern of protein tyrosine phosphorylation was observed in the lysates of T cells prepared from seven healthy young adults and eight healthy elderly human subjects. Five different high molecular mass proteins (75, 115, 120, 140 and 170 kDa) were consistently tyrosine phosphorylated in all of the donors from both age groups and peaked between 3 and 10 min. Tyrosine phosphorylation of the above substrates was observed in both CD4 and CD8 subsets. When compared for individual donors from both age groups, variations in the T cell response with regard to net tyrosine phosphorylation for all the substrates was observed. However, the mitogen induced level of tyrosine phosphorylation of only p75 was found to be significantly lower in unfractionated T cells as well as CD4 and CD8 subsets of older subjects than that of young subjects. Using immunoblotting, p75 was identified as ZAP-70, a member of the syk family of protein tyrosine kinases. Understanding of the biochemical basis of the reduced level of tyrosine phosphorylation of ZAP-70 will be helpful in delineating the molecular basis of age-associated impairment of T cell activation.

Adult↗