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K Iqbal

Publications and source records attributed to K Iqbal.

At least 73 records · Page 4Linked to original sources
PubMed1997

Abnormal phosphorylation of tau and the mechanism of Alzheimer neurofibrillary degeneration: sequestration of microtubule-associated proteins 1 and 2 and the disassembly of microtubules by the abnormal tau.

The microtubule-associated protein (MAP) tau is abnormally hyperphosphorylated in Alzheimer disease and accumulates in neurons undergoing neurofibrillary degeneration. In the present study, the associations of the Alzheimer-hyperphosphorylated tau (AD P-tau) with the high molecular weight MAPs (HMW-MAPs) MAP1 and MAP2 were investigated. The AD P-tau was found to aggregate with MAP1 and MAP2 in solution. The association of AD P-tau to the MAPs resulted in inhibition of MAP-promoted microtubule assembly. However, unlike the coaggregation of AD P-tau and normal tau, the association between AD P-tau and the HMW-MAPs did not result in the formation of filaments/tangles. The affinity of the tau-AD P-tau association was higher than that of HMW-MAPs-AD P-tau because normal tau inhibited the latter binding. The association between AD P-tau and the HMW-MAPs also appeared to occur in situ because these proteins cosedimented from the Alzheimer brain extracts, and, in the sediment, the levels of the HMW-MAPs correlated with the levels of AD P-tau. These studies suggested that the abnormally phosphorylated tau can sequester both normal tau and HMW-MAPs and disassemble microtubules but, under physiological conditions, can form tangles of filaments only from tau.

Alzheimer Disease↗
PubMed1997

Alzheimer disease hyperphosphorylated tau aggregates hydrophobically.

The chemical interaction that condenses the hyperphosphorylated protein tau in Alzheimer's disease (AD P-tau) into neurofibrillary tangles and cripples synaptic transmission remains unknown. Only beta-sheet, positive ion salt bridges between phosphates, and hydrophobic association can create tangles of just AD P-tau. We have correlated transmission electron microscope (TEM) images of tau aggregation with different percentages of beta-sheet in aqueous suspensions of tau while using buffers that block dispositive or tripositive ionic bridges between intermolecular phosphates. Circular dichroism (CD) studies were performed at different temperatures from 5-85 degrees C using AD P-tau, AD P-tau dephosphorylated with hydrofluoric acid (HF AD P-tau) or alkaline phosphatase (AP AD P-tau), and recombinant human tau with 3-repeats and two amino terminal inserts (R-39) and using bovine tau (B tau) isolated without heat or acid treatment. Secondary structure was estimated from CD spectra at 5 degrees C using the Lincomb algorithm. Each preparation except one demonstrated an inverse temperature transition, Ti, in the CD at 197 nm. No correlation was found between beta-sheet content and aggregation, leaving only hydrophobic interaction as the remaining possibility. Thirteen of 21 possible phosphorylation sites in AD P-tau lie adjacent to positive residues in tau's primary structure. Occupation of five to nine phosphate sites on AD P-tau appears sufficient to reduce or neutralize tau's basic character. AD P-tau's hydrophobic character is indicated by its low inverse temperature transition, Ti. The Ti for AD P-tau was 24.5 degrees C or 28 degrees C, whereas for B tau with three phosphates it was 32 degrees C, for unphosphorylated tau R-39 it was 38 degrees C, and for dephosphorylated HF AD P-tau it was 37.5 degrees C. The hydrophobic protein elastin and its analogs coalesce and precipitate at their Ti of 24-29 degrees C, well below body temperature. We hypothesize that AD P-tau causes tangle accumulation by this mechanism.

Alkaline Phosphatase↗
PubMed1997

Elevated protein levels of protein phosphatases PP-2A and PP-2B in astrocytes of Alzheimer's disease temporal cortex.

Previous studies have shown that activities of the protein phosphatases PP-2A and PP-2B towards the microtubule associated protein tau are reduced in Alzheimer's disease (AD) frontal cortex (Gong et al., 1993, 1995), suggesting that PP-2A and PP-2B are involved in the hyperphosphorylation of tau in AD. Most recently, we found that protein levels of PP-2A and PP-2B are elevated in postsynaptic supernatant (S2) fractions prepared from AD temporal cortex, and that the activities of these enzymes were not significantly different between AD and control cases (Pei et al., in press). In the present study, we found that astroglia positive for PP-2A and PP-2B immunoreactivities were greater in numbers in AD medial temporal cortex, compared to controls. GFAP levels, as determined by indirect ELISA, were approximately 1.5 times greater in the P1 (500 x g) fraction from AD temporal cortex, compared to controls. GFAP levels in the P1 fraction showed significant correlations with PP-2A and PP-2B levels in the postsynaptic S2 (20,000 x g) fraction from the same brains. These results suggest that astrogliosis probably accounts for the increased levels of PP-2A and PP-2B in the S2 fraction in AD brain and that the levels of these enzymes per neuron are likely to be decreased.

Aged↗
PubMed1997

Elevated levels of tau and ubiquitin in brain and cerebrospinal fluid in Alzheimer's disease.

Alzheimer's disease (AD) is histopathologically characterized by the presence of numerous neurons with neurofibrillary tangles of paired helical filaments (PHF) in the neocortex, particularly the hippocampus. The major protein subunit of PHF, which also accumulate as neuropil threads and dystrophic neurites of the neuritic (senile) plaques, is the microtubule-associated protein tau. tau in AD brain is abnormally hyperphosphorylated. In addition to hyperphosphorylation, some of the AD tau is also ubiquitinated. The level of tau in AD brain is elevated severalfold, and this increase is in the form of the abnormally phospholylated tau. The levels of conjugated ubiquitin are also increased severalfold in AD brain. The cerebrospinal fluid (CSF) levels of both tau and ubiquitin are elevated in AD. However, some of the normal elderly and non-AD neurological control cases also show elevated levels of these two proteins in the CSF. At present, it is not clear whether the overlap in CSF tau or ubiquitin values between AD and the control groups is due to some unknown heterogeneity of AD, inclusion of presymptomatic AD cases in the control group, or lack of specificity of the CSF elevation of these markers to AD. Thus, until the results of prospective studies on many patients with AD and normal controls become available, the CSF tau and ubiquitin levels can be used only as additional tools in the psychometric and clinical diagnosis of AD.

Aged↗
PubMed1997

Protein kinase C and calcium/calmodulin-dependent protein kinase II phosphorylate three-repeat and four-repeat tau isoforms at different rates.

All six isoforms of the microtubule-associated protein tau are present in hyperphosphorylated states in the brains of patients with Alzheimer's disease (AD). It is presently unclear how such hyperphosphorylation of tau is controlled. In a previous study (Singh et al. Arch Biochem Biophys 328: 43-50, 1996) we have shown that three-repeat taus containing two N-terminal inserts were phosphorylated to higher levels and at different sites compared to those either lacking or containing only one such insert. We have extended these observations in this study by comparing the phosphorylation of tau isoforms containing three-repeats (tau 3, tau 3 L) and four-repeats (tau 4, tau 4 L). In the absence of N-terminal inserts in tau structure (tau 3, tau 4) both CaM kinase II and C-kinase phosphorylated four-repeat tau (tau 4) to a higher extent than three-repeat tau (tau 3). When two N-terminal inserts are present in tau structure (tau 3 L, tau 4 L), then three-repeat tau (tau 3 L) is phosphorylated to a higher extent than four-repeat tau (tau 4 L) by these kinases. CK-1 and GSK-3 phosphorylated each of the above pairs of three-repeat and four-repeat taus to the same extents. However, after an initial prephosphorylation of the taus by CaM kinase II, GSK-3 differentially phosphorylated three-repeat and four-repeat taus. Under these conditions thr 231, ser 235, ser 396, and ser 404 were phosphorylated to greater extents in four-repeat tau (tau 4) compared to three-repeat tau (tau 3) in the absence of N-terminal inserts. In the presence of such inserts these sites were phosphorylated to greater extents in three-repeat (tau 3 L) compared to four-repeat (tau 4 L) tau. Our results indicate that the extents to which tau isoforms are phosphorylated in normal and AD brain depends on (a) the number of repeats (3 or 4), (b) the number of N-terminal inserts (0, 1, or 2), and (c) the initial phosphorylation state of tau.

Calcium-Calmodulin-Dependent Protein Kinase Type 2↗
PubMed1997

Potentiation of GSK-3-catalyzed Alzheimer-like phosphorylation of human tau by cdk5.

Tau protein from Alzheimer disease (AD) brain is hyperphosphorylated by both proline-dependent protein kinases (PDPKs) and non-PDPKs. It is presently unclear how PDPKs and non-PDPKs interact in tau hyperphosphorylation. Previously we have shown that non-PDPKs can positively modulate the activity of a PDPK (GSK-3) in tau phosphorylation (Singh et al. (1995) FEBS Lett. 358, 267-272). In this study we have investigated whether (A) non-PDPKs can also modulate the activity of the PDPK, cdk5, (B) a PDPK can modulate the activities of another PDPK, as well as non-PDPKs. We found that, like GSK-3, the activity of cdk5 is stimulated if tau were first prephosphorylated by any of several non-PDPKs (A-kinase, C-kinase, CK-1, CaM-kinase II). Prephosphorylation of tau by cdk5 stimulated both the rate and extent of a subsequent phosphorylation catalyzed by GSK-3. Under these conditions thr 231 phosphorylation was especially enhanced (9-fold). No significant stimulation of phosphorylation was observed when the order of these kinases was reversed (i.e. GSK-3 followed by cdk5). By contrast, prephosphorylation of tau by cdk5 served to inhibit subsequent phosphorylation catalyzed by C-kinase and CK-1, but not by A-kinase or CaM-kinase II. Our results suggest that in tau hyperphosphorylation in AD brain, cdk5-catalyzed phosphorylation may serve to upregulate the activity of GSK-3 and down-regulate the activities of C-kinase and CK-1.

Alzheimer Disease↗
PubMed1997

Distribution, levels, and activity of glycogen synthase kinase-3 in the Alzheimer disease brain.

A number of studies have implicated a proline-directed protein kinase, glycogen synthase kinase-3 (GSK-3) in the hyperphosphorylation of tau in Alzheimer's disease (AD). Toward understanding the role of GSK-3 in the abnormal hyperphosphorylation of tau in AD we have found that GSK-3 is prominently present in neuronal cell bodies and their processes and co-localizes with neurofibrillary changes in AD brain. Furthermore, the levels of GSK-3 as determined by indirect ELISA are approximately 50% increased in the postsynaptosomal supernatant from AD brains as compared to the controls. However, no increase in GSK-3 enzyme activity was detected. In AD brain, with its reduced phosphatase activity, even normal levels of GSK-3 activity might be sufficient for the hyperphosphorylation of tau.

Alzheimer Disease↗
PubMed1996

Inhibition of protein phosphatase-2B (calcineurin) activity towards Alzheimer abnormally phosphorylated tau by neuroleptics.

Abnormally hyperphosphorylated tau is the major protein component of neurofibrillary tangles, the characteristic lesion of Alzheimer's disease (AD). Protein phosphatases (PP) type 1 (PP-1), type 2A (PP-2A) and type 2B (PP-2B) appear to be involved in the regulation of tau phosphorylation. The incidence of neurofibrillary tangles is higher in brains of schizophrenic patients treated with neuroleptics than in those without this treatment. We have found that the commonly used neuroleptics chlorpromazine, trifluoperazine and clozapine inhibit PP-2B but not PP-1 or PP-2A activity towards [32P]phosphorylase kinase as a substrate. When AD abnormally hyperphosphorylated tau is used as a substrate, PP-2B activity is inhibited by trifluoperazine > chlorpromazine > clozapine. Using phosphorylation-dependent monoclonal antibodies, tau-1, AT8 and PHF-1, we have found that the dephosphorylation of the abnormal tau by PP-2B is inhibited at all the sites recognized by these antibodies. The IC50 of the inhibition of dephosphorylation at tau-1 site is approximately 20 microM for trifluoperazine and approximately 120 microM for chlorpromazine. These two neuroleptics inhibit tau dephosphorylation by PP-2B through antagonizing calmodulin as well as directly interacting with PP-2B. The inhibition of the dephosphorylation of abnormally hyperphosphorylated tau by neuroleptics raises an intriguing possibility that the chronic use of these drugs might contribute to neurofibrillary degeneration in schizophrenic and AD patients.

Alzheimer Disease↗
PubMed1996

Calcium/calmodulin-dependent protein kinase II phosphorylates tau at Ser-262 but only partially inhibits its binding to microtubules.

PHF-tau, which is phosphorylated at 10 Ser/Thr-Pro and 11 non-Ser/Thr-Pro sites, is unable to promote microtubule assembly. Phosphorylation of the non-Ser/Thr-Pro site, Ser-262, is reported to be primarily responsible for this. The identities of kinase(s) responsible for Ser-262 phosphorylation are still to be clarified. In this study we have used the monoclonal antibody 12E8, which recognizes P-Ser-262 and P-Ser-356 on tau, to survey different kinases for their abilities to phosphorylate Ser-262 on human tau 3L (tau410). In decreasing order of effectiveness we found that Ser-262 and Ser-356 phosphorylation can be catalyzed by CaM kinase II >> C-kinase >> GSK-3 approximately = A-kinase >> CK-1. CaM kinase II and C-kinase were shown to phosphorylate both Ser-262 and Ser-356. The binding of tau to taxol-stabilized microtubules was decreased by 35 and 42% after phosphorylation by CaM kinase II and C-kinase, respectively. Of the fraction of tau that bound to microtubules, about 50% was phosphorylated at Ser-262 and Ser-356. These results suggest that Ser-262 and Ser-356 are very good substrates for CaM kinase II but their phosphorylations are not sufficient to achieve maximal inhibition of tau binding to microtubules.

Binding Sites↗
PubMed1996

Differential phosphorylation of human tau isoforms containing three repeats by several protein kinases.

The paired helical filaments (PHF) found in the brain of patients with Alzheimer disease (AD) are composed primarily of the microtubule-associated protein tau. Six isoforms of tau have been recognized and all are present in a hyperphosphorylated state in PHF. It is not known whether all tau isoforms serve equally well as substrates for various kinases. In this study we have compared the phosphorylation of human tau isoforms containing three microtubule-binding repeats and zero (tau 3), one (tau 3S), or two (tau 3L) N-terminal inserts. Four kinases (A-kinase, CK-1, CaM kinase II, GSK-3) were used for this purpose. With A-kinase, CK-1, and CaM kinase II the extent of phosphorylation was tau 3L > tau 3S > tau 3. With GSK-3 it was tau 3L approximately = tau 3S > tau 3. Tau 3 was a poor substrate for either CaM kinase II or CK-1, 32P incorporation being only 5 and 11%, respectively, of that observed by these kinases when tau 3L was the substrate. After prephosphorylation of the three tau isoforms by A-kinase, a subsequent phosphorylation by GSK-3 was stimulated several fold over tau that was not prephosphorylated. Under these conditions the extent of 32P incorporation was tau 3L > tau 3S > tau 3. Both CK-1 and GSK-3 phosphorylated ser 396 more rapidly in tau 3L compared to tau 3 or tau 3S. Our results suggest that (1) the presence of N-terminal inserts in tau isoforms are important structural determinants that modulate the specificity of several kinases; (2) the different tau isoforms may be present at different states of phosphorylation in PHF.

Calcium-Calmodulin-Dependent Protein Kinases↗
PubMed1996

Non-proline-dependent protein kinases phosphorylate several sites found in tau from Alzheimer disease brain.

Of 21 phosphorylation sites identified in PHF-tau 11 are on ser/thr-X motifs and are probably phosphorylated by non-proline-dependent protein kinases (non-PDPKs). The identities of the non-PDPKs and how they interact to hyperphosphorylate PHF-tau are still unclear. In a previous study we have shown that the rate of phosphorylation of human tau 39 by a PDPK (GSK-3) was increased several fold if tau were first prephosphorylated by non-PDPKs (Singh et al., FEBS Lett 358: 267-272, 1995). In this study we have examined how the specificity of a non-PDPK for different sites on human tau 39 is modulated when tau is prephosphorylated by other non-PDPKs (A-kinase, C-kinase, CK-1, CaM kinase II) as well as a PDPK (GSK-3). We found that the rate of phosphorylation of tau 39 by a non-PDPK can be stimulated if tau were first prephosphorylated by other non-PDPKs. Of the four non-PDPKs only CK-1 can phosphorylate sites (thr 231, ser 396, ser 404) known to be present in PHF-tau. Further, these sites were phosphorylated more rapidly and to a greater extent by CK-1 if tau 39 were first prephosphorylated by A-kinase, CaM kinase II or GSK-3. These results suggest that the site specificities of the non-PDPKs that participate in PHF-tau hyperphosphorylation can be modulated at the substrate level by the phosphorylation state of tau.

Alzheimer Disease↗
PubMed1996

Molecular mechanism of Alzheimer's neurofibrillary degeneration and therapeutic intervention.

Microtubule-associated protein tau is abnormally hyperphosphorylated in the brain of patients with Alzheimer's disease (AD) and the abnormal tau is the major protein subunit of paired helical filaments (PHF). The abnormal phosphorylation of tau probably precedes its polymerization into PHF. The abnormal tau does not bind to tubulin, but competes with tubulin in binding to normal tau and thereby inhibits the assembly of microtubules in the affected neurons. The abnormal tau can be dephosphorylated enzymatically and by this way its microtubule assembly promoting activity can be restored. The activities of protein phosphatases might be decreased in the affected neurons in AD brain, allowing the abnormal hyperphosphorylation of tau. Neurofibrillary degeneration can probably be inhibited by increasing the activities of protein phosphatases in the brain of patients with Alzheimer's disease.

Alzheimer Disease↗
PubMed1996

Restoration of biological activity of Alzheimer abnormally phosphorylated tau by dephosphorylation with protein phosphatase-2A, -2B and -1.

Microtubule associated protein tau promotes the assembly of microtubules by binding to microtubules and stabilizing their structure. In Alzheimer disease brain, tau is abnormally hyperphosphorylated and the altered tau is unable to promote the in vitro assembly of microtubules. In the present study, we found that dephosphorylation of abnormally phosphorylated tau by protein phosphatase-2A1, -2B or -1 restored its biological activity both in the nucleation and in the assembly of microtubules. Both the amount of phosphate released and the rate of restoration of microtubule assembly promoting activity of the abnormal tau were greater on dephosphorylation by protein phosphatase-2A1 than -2B or -1. During 90 min incubation at 37 degrees C protein phosphatase-2A1, -2B and -1 released respectively approximately 57%, approximately 36% and approximately 30% of tau phosphate. Association of the restoration of the biological activity of the abnormal tau dephosphorylated by different phosphatases and the immunochemical identification of the dephosphorylated sites revealed that Ser-235 is not critical in tau function, and that the Thr-231 is probably involved in the regulation of the nucleation and not the assembly of microtubules. These studies indicate that the phosphorylation of tau in situ might be regulated by protein phosphatase-2A, -2B and -1 and activation of these enzyme activities might arrest the Alzheimer neurofibrillary degeneration.

Alkaline Phosphatase↗
PubMed1996

Alzheimer's disease hyperphosphorylated tau sequesters normal tau into tangles of filaments and disassembles microtubules.

Microtubule-associated protein tau becomes abnormally hyperphosphorylated in Alzheimer's disease (AD) and accumulates as tangles of paired helical filaments in neurons undergoing degeneration. We now show that in solution normal tau associates with the AD hyperphosphorylated tau (AD P-tau) in a nonsaturable fashion, forming large tangles of filaments 3.3 +/- 0.7 nm in diameter. These tangles, which are not detected in identically treated normal tau or AD P-tau alone, are made up of filaments several microns in length and are labeled with tau antibodies. Dephosphorylation with alkaline phosphatase abolishes the ability of AD P-tau to aggregate with normal tau and prevents tangle formation. AD P-tau disassembles microtubules assembled from normal tau and tubulin. These data provide insight into how the hyperphosphorylation of tau might lead to the formation of the neurofibrillary tangles and the degeneration of the affected neurons in AD.

Alzheimer Disease↗
PubMed1996

Glycosylation of microtubule-associated protein tau: an abnormal posttranslational modification in Alzheimer's disease.

Alzheimer's disease (AD) is characterized by the presence of numerous neurons with neurofibrillary tangles of paired helical filaments (PHFs). The microtubule-associated protein tau in abnormally hyperphosphorylated form is the major protein subunit of the PHF. We now show that PHF tangles isolated from AD brains are glycosylated, whereas no glycan is detected in normal tau. Deglycosylation of PHF tangles by endoglycosidase F/N-glycosidase F converts them into bundles of straight filaments 2.5 +/- 0.5 nm in diameter, similar to those generated by the interaction of normal tau and abnormally hyperphosphorylated tau (AD P-tau). Deglycosylation plus dephosphorylation, but not deglycosylation alone, of AD P-tau and tau from PHF tangles restores their microtubule polymerization activity. Dephosphorylation of deglycosylated PHF tangles results in increased tau release. Thus, although the abnormal phosphorylation might promote aggregation of tau and inhibition of the assembly of microtubules, glycosylation appears to be responsible for the maintenance of the PHF structure.

Alzheimer Disease↗
PubMed1996

Stomal recurrence after total laryngectomy.

Radical laryngeal surgery was performed in 71 patients of stage III and IV laryngeal carcinoma. They were followed-up for the stomal recurrence. Stomal recurrence occurred in 5 (7%) patients, four of these had cervical nodal metastasis. The average time interval between laryngectomy and the stomal recurrence was 19 months. All the patients expired and the mean length of survival after the recurrence was 5.3 months.

Adult↗
PubMed1996

Laryngeal involvement in pulmonary tuberculosis.

Fifty-one patients with established pulmonary tuberculosis underwent clinical evaluation and endoscopic examination of the larynx to determine the manifestations of laryngeal involvement. There were 46 males and 5 females (mean age 38 years). Fever, cough and haemoptysis were the prime pulmonary complaints while hoarseness, weak voice and episodic dyspnoea were the main laryngeal symptoms. Sites of laryngeal lesions included true vocal cords, arytenoids and false vocal cords. Oedema, pallor, ulcers, vocal cord immobility and thickening were the main laryngeal lesions observed. Twenty-four (47%) cases showed morphological changes in the larynx. The presentation pattern was consistent with the classical description and predilection for laryngeal involvement was not demonstrated.

Adult↗