{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,25]],"date-time":"2026-07-25T15:04:20Z","timestamp":1784991860514,"version":"3.55.0"},"reference-count":27,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5333","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1997,9,19]]},"abstract":"<jats:p>Classical late-infantile neuronal ceroid lipofuscinosis (LINCL) is a fatal neurodegenerative disease whose defective gene has remained elusive. A molecular basis for LINCL was determined with an approach applicable to other lysosomal storage diseases. When the mannose 6-phosphate modification of newly synthesized lysosomal enzymes was used as an affinity marker, a single protein was identified that is absent in LINCL. Sequence comparisons suggest that this protein is a pepstatin-insensitive lysosomal peptidase, and a corresponding enzymatic activity was deficient in LINCL autopsy specimens. Mutations in the gene encoding this protein were identified in LINCL patients but not in normal controls.<\/jats:p>","DOI":"10.1126\/science.277.5333.1802","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T05:37:20Z","timestamp":1027748240000},"page":"1802-1805","source":"Crossref","is-referenced-by-count":483,"title":["Association of Mutations in a Lysosomal Protein with Classical Late-Infantile Neuronal Ceroid Lipofuscinosis"],"prefix":"10.1126","volume":"277","author":[{"given":"David E.","family":"Sleat","sequence":"first","affiliation":[{"name":"D. E. Sleat, H. Lackland, C.-G. Liu, I. Sohar, P. Lobel, Center for Advanced Biotechnology and Medicine, Piscataway, NJ 08854, USA and Department of Pharmacology, Robert Wood Johnson Medical School\u2013University of Medicine and Dentistry of New Jersey, NJ 08854, USA."},{"name":"R. J. Donnelly, Department of Lab Medicine and Pathology, New Jersey Medical School\u2013University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA."},{"name":"R. K. Pullarkat, New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY 10314, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert J.","family":"Donnelly","sequence":"additional","affiliation":[{"name":"D. E. Sleat, H. Lackland, C.-G. Liu, I. Sohar, P. Lobel, Center for Advanced Biotechnology and Medicine, Piscataway, NJ 08854, USA and Department of Pharmacology, Robert Wood Johnson Medical School\u2013University of Medicine and Dentistry of New Jersey, NJ 08854, USA."},{"name":"R. J. Donnelly, Department of Lab Medicine and Pathology, New Jersey Medical School\u2013University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA."},{"name":"R. K. Pullarkat, New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY 10314, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henry","family":"Lackland","sequence":"additional","affiliation":[{"name":"D. E. Sleat, H. Lackland, C.-G. Liu, I. Sohar, P. Lobel, Center for Advanced Biotechnology and Medicine, Piscataway, NJ 08854, USA and Department of Pharmacology, Robert Wood Johnson Medical School\u2013University of Medicine and Dentistry of New Jersey, NJ 08854, USA."},{"name":"R. J. Donnelly, Department of Lab Medicine and Pathology, New Jersey Medical School\u2013University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA."},{"name":"R. K. Pullarkat, New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY 10314, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chang-Gong","family":"Liu","sequence":"additional","affiliation":[{"name":"D. E. Sleat, H. Lackland, C.-G. Liu, I. Sohar, P. Lobel, Center for Advanced Biotechnology and Medicine, Piscataway, NJ 08854, USA and Department of Pharmacology, Robert Wood Johnson Medical School\u2013University of Medicine and Dentistry of New Jersey, NJ 08854, USA."},{"name":"R. J. Donnelly, Department of Lab Medicine and Pathology, New Jersey Medical School\u2013University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA."},{"name":"R. K. Pullarkat, New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY 10314, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Istvan","family":"Sohar","sequence":"additional","affiliation":[{"name":"D. E. Sleat, H. Lackland, C.-G. Liu, I. Sohar, P. Lobel, Center for Advanced Biotechnology and Medicine, Piscataway, NJ 08854, USA and Department of Pharmacology, Robert Wood Johnson Medical School\u2013University of Medicine and Dentistry of New Jersey, NJ 08854, USA."},{"name":"R. J. Donnelly, Department of Lab Medicine and Pathology, New Jersey Medical School\u2013University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA."},{"name":"R. K. Pullarkat, New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY 10314, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raju K.","family":"Pullarkat","sequence":"additional","affiliation":[{"name":"D. E. Sleat, H. Lackland, C.-G. Liu, I. Sohar, P. Lobel, Center for Advanced Biotechnology and Medicine, Piscataway, NJ 08854, USA and Department of Pharmacology, Robert Wood Johnson Medical School\u2013University of Medicine and Dentistry of New Jersey, NJ 08854, USA."},{"name":"R. J. Donnelly, Department of Lab Medicine and Pathology, New Jersey Medical School\u2013University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA."},{"name":"R. K. Pullarkat, New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY 10314, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Lobel","sequence":"additional","affiliation":[{"name":"D. E. Sleat, H. Lackland, C.-G. Liu, I. Sohar, P. Lobel, Center for Advanced Biotechnology and Medicine, Piscataway, NJ 08854, USA and Department of Pharmacology, Robert Wood Johnson Medical School\u2013University of Medicine and Dentistry of New Jersey, NJ 08854, USA."},{"name":"R. J. Donnelly, Department of Lab Medicine and Pathology, New Jersey Medical School\u2013University of Medicine and Dentistry of New Jersey, Newark, NJ 07103, USA."},{"name":"R. K. Pullarkat, New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY 10314, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","unstructured":"R.-M. Boustany in Neurodystrophies and Neurolipidoses vol. 66 of Handbook of Clinical Neurology H. W. Moser Ed. (Elsevier Amsterdam 1996) pp. 671\u2013700."},{"key":"e_1_3_1_3_2","doi-asserted-by":"crossref","unstructured":"J. A. Rider G. Dawson A. N. Siakotos Am. J. Med. Genet. 42 519 24 (1992).","DOI":"10.1002\/ajmg.1320420419"},{"key":"e_1_3_1_4_2","doi-asserted-by":"crossref","first-page":"591","DOI":"10.1093\/hmg\/6.4.591","volume":"6","author":"Sharp J. D.","year":"1997","unstructured":"Sharp J. D., et al., Hum. Mol. Genet. 6, 591 (1997).","journal-title":"Hum. Mol. Genet."},{"key":"e_1_3_1_5_2","doi-asserted-by":"crossref","first-page":"584","DOI":"10.1038\/376584a0","volume":"376","author":"Vesa J.","year":"1995","unstructured":"Vesa J., et al., Nature 376, 584 (1995).","journal-title":"Nature"},{"key":"e_1_3_1_6_2","doi-asserted-by":"crossref","unstructured":"The International Battens Disease Consortium Cell 82 949 (1995).","DOI":"10.1016\/0092-8674(95)90274-0"},{"key":"e_1_3_1_7_2","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1126\/science.6505679","volume":"226","author":"Ivy G. O.","year":"1984","unstructured":"Ivy G. O., Schottler F., Wenzel J., Baudry M., Lynch G., Science 226, 985 (1984).","journal-title":"Science"},{"key":"e_1_3_1_8_2","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1002\/ajmg.1320420427","volume":"42","author":"Ivy G. O.","year":"1992","unstructured":"Ivy G. O., Am. J. Med. Genet. 42, 555 (1992).","journal-title":"Am. J. Med. Genet."},{"key":"e_1_3_1_9_2","unstructured":"S. Kornfeld and W. S. Sly The Metabolic and Molecular Bases of Inherited Disease C. R. Sciver A. L. Beaudet W. S. Sly D. Valle Eds. (McGraw-Hill New York 1995) vol. 2."},{"key":"e_1_3_1_10_2","doi-asserted-by":"crossref","first-page":"19191","DOI":"10.1074\/jbc.271.32.19191","volume":"271","author":"Sleat D. E.","year":"1996","unstructured":"Sleat D. E., Sohar I., Lackland H., Majercak J., Lobel P., J. Biol. Chem. 271, 19191 (1996).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_11_2","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1042\/bj3240033","volume":"324","author":"Sleat D. E.","year":"1997","unstructured":"Sleat D. E., Kraus S. R., Sohar I., Lackland H., Lobel P., Biochem. J. 324, 33 (1997).","journal-title":"Biochem. J."},{"key":"e_1_3_1_12_2","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1006\/abio.1993.1096","volume":"209","author":"Valenzano K. J.","year":"1993","unstructured":"Valenzano K. J., Kallay L. M., Lobel P., Anal. Biochem. 209, 156 (1993).","journal-title":"Anal. Biochem."},{"key":"e_1_3_1_13_2","unstructured":"Human EST cDNA clones zo55e03 EST37588 and zo35g10 were sequenced in their entirety. Clones zs52e09 and zr50co6 were partially sequenced and appear to contain cloning artifacts."},{"key":"e_1_3_1_14_2","unstructured":"The 5\u2032 end of the human cDNA was obtained by two rounds of PCR amplification of the CLN2 candidate from a human cortex cDNA library (Stratagene) by means of two different gene-specific primers and a single vector-specific primer. In the first round of PCR the T3 promoter primer was used with either gene-specific primer NR1 (5\u2032-GTGATCACAGAATGGCACTT) or NR2 (5\u2032-AACATGGGTTTCCGTAGGTC). In the second round of PCR with the products from the first amplification the T3 promoter primer and NR4 (5\u2032-CTTCCTCAGGGTCCGCACGG) were used."},{"key":"e_1_3_1_15_2","unstructured":"GenBank accession number ."},{"key":"e_1_3_1_16_2","unstructured":"Three lines of evidence give corroborative results for an unequivocal localization. (i) There is a nearly perfect match between nt 34 to 104 of the CLN2 cDNA candidate and GenBank accession number which represents a PCR-amplified fragment of a flow-sorted chromosome 11\u2013specific cosmid clone. (The 317-nt  also contains sequence of flanking introns.) (ii) There is a perfect 505-nt match between the 3\u2032 end of the CLN2 cDNA (nt 2979 to 3483) and the 5\u2032 end (nt 1 to 505) of GenBank accession number .  consists of 2605 nt that encompass the human TATA-binding protein associated factor II 30 (TAF II 30) gene. The TAF II 30 transcription start site is at  nt 1060 and most of the promoter elements are downstream of  nt 860 and thus do not overlap with the 3\u2032 end of the large CLN2 candidate transcript. Thus the CLN2 candidate gene and the TAF II 30 gene are physically adjacent. The TAF II 30 gene was mapped to chromosome 11p15.2-p15.5 by means of in situ hybridization ["},{"key":"e_1_3_1_16_3","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1006\/geno.1995.1243","volume":"29","author":"Scheer E.","year":"1995","unstructured":"Scheer E., Mattei M. G., Jacq X., Chambon P., Tora L., Genomics 29, 269 (1995);","journal-title":"Genomics"},{"key":"e_1_3_1_16_4","unstructured":"]. (iii) Three sequences (accession numbers and ) representing a cosmid clone have strong matches ( P < 10 \u2212 31 ) to nt 2817 to 3264 of the CLN2 candidate cDNA. The cosmid clone maps to chromosome 11p15. Taken together these results indicate that the CLN2 candidate is localized to chromosome 11p15."},{"key":"e_1_3_1_17_2","unstructured":"D. E. Sleat et al. unpublished data."},{"key":"e_1_3_1_18_2","unstructured":"CLN2 was analyzed in patient DNA extracted from cell lines by means of overlapping M13 forward- and reverse-tailed primer pairs. Each pair amplified an exon and flanking intronic sequences and the resulting products were sequenced with dye-labeled \u221221M13 primer. For patients the sequence of fragments that mismatched with the consensus sequence was then confirmed by sequencing with the M13 reverse primer. Each fragment containing a mutation in both patients and relatives was then independently reamplified and sequenced on both strands to confirm that the observed heterogeneities were not artifacts of PCR amplification. Primer pairs that detected mutations in patient DNA were SF3 (5\u2032-TGTAAAACGACGGCCAGTCAGACCTTCCAGTAGGGACC)\/SR3 (5\u2032-CAGGAAACAGCTATGACCCTGTATCCCACACAAGAGAT) and SF0A (5\u2032-TGTAAAACGACGGCCAGTTAGATGCCATTGGGGACTGG)\/SR0A (5\u2032-CAGGAAACAGCTATGACCGTCATGGAAATACTGCTCCA). PCR from 1-\u03bcg samples of patient DNA with Vent DNA polymerase (New England Biolabs Beverly MA) was done under the following cycle conditions: 94\u00b0C for 3 min followed by 10 cycles of 94\u00b0C for 1 min 50\u00b0C for 1 min and 72\u00b0C for 1 min followed by 30 cycles of 94\u00b0C for 1 min 65\u00b0C for 1 min and 72\u00b0C for 1 min with a final incubation for 10 min at 72\u00b0C. Products were purified by means of Qiaquick spin columns (Quiagen Chatsworth CA) and cycle-sequenced with AmpliTaq DNA polymerase (Roche Molecular Systems Alameda CA) and ABI Prism dye-labeled primers (Perkin Elmer Foster City CA) on an ABI 373 automated sequencer."},{"key":"e_1_3_1_19_2","unstructured":"A BLAST search of the SwissProt database with the conceptually translated CLN2 candidate gave a highly significant match with PsCP: P = 1.9 \u00d7 10 \u2212 11 ; the Dayhoff comparison score is >8 SDs above the mean (ALIGN program relative to 200 comparisons of scrambled sequences); and pairwise comparison with GCG Bestfit yields identity and similarity scores of 25 and 46% respectively. PsCP is related (52% identical 66% similar) to XaCP. XaCP is not detected in a BLAST search with the CLN2 candidate but in pairwise comparisons the Dayhoff comparison score is >2.7 SDs above the mean and the identity and similarity scores are 24 and 48% respectively."},{"key":"e_1_3_1_20_2","doi-asserted-by":"crossref","first-page":"26518","DOI":"10.1016\/S0021-9258(18)47225-0","volume":"269","author":"Oda K.","year":"1994","unstructured":"Oda K., Takahashi T., Tokuda Y., Shibano Y., Takahashi S., J. Biol. Chem. 269, 26518 (1994).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_21_2","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1093\/oxfordjournals.jbchem.a021451","volume":"120","author":"Oda K.","year":"1996","unstructured":"Oda K., et al., J. Biochem. 120, 564 (1996).","journal-title":"J. Biochem."},{"key":"e_1_3_1_22_2","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1002\/ajmg.1320420428","volume":"42","author":"Palmer D. N.","year":"1992","unstructured":"Palmer D. N., Fearnley I. M., Medd S. M., Am. J. Med. Genet. 42, 561 (1992).","journal-title":"Am. J. Med. Genet."},{"key":"e_1_3_1_23_2","doi-asserted-by":"crossref","first-page":"5736","DOI":"10.1016\/S0021-9258(18)83610-9","volume":"264","author":"Palmer D. N.","year":"1989","unstructured":"Palmer D. N., et al., J. Biol. Chem. 264, 5736 (1989).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_24_2","doi-asserted-by":"crossref","first-page":"1677","DOI":"10.1046\/j.1471-4159.1996.67041677.x","volume":"67","author":"Ezaki J.","year":"1996","unstructured":"Ezaki J., Wolfe L. S., Kominami E., J. Neurochem. 67, 1677 (1996).","journal-title":"J. Neurochem."},{"key":"e_1_3_1_25_2","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1016\/0003-9861(74)90490-1","volume":"163","author":"De Bernardo S.","year":"1974","unstructured":"De Bernardo S., et al., Arch. Biochem. Biophys. 163, 390 (1974).","journal-title":"Arch. Biochem. Biophys."},{"key":"e_1_3_1_26_2","unstructured":"We thank P. S. Pullarkat for technical assistance and A. B. Rabson and S. A. Moodie for providing cDNA libraries. Supported by NIH grant DK45992 and a gift from Pfizer (P.L.) and by NIH grant NS30147 (R.K.P.)."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.277.5333.1802","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T00:00:59Z","timestamp":1705104059000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.277.5333.1802"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1997,9,19]]},"references-count":27,"journal-issue":{"issue":"5333","published-print":{"date-parts":[[1997,9,19]]}},"alternative-id":["10.1126\/science.277.5333.1802"],"URL":"https:\/\/doi.org\/10.1126\/science.277.5333.1802","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1997,9,19]]}}}