Tumor Endothelial Marker Report: TEM14 (cystatin - undetermined subtype)

	TEM domain structure

	TEM Sequence Analysis

name	from	to	source/E	description
SignalP	1	21
cystatin	32	137	Pfam, E = 9.4e-40	The cystatin domain contains three conserved cystatins motifs: G32; two motifs involved in cysteine proteinase inhibition: Q77-G80, VPW127; and the cysteines 94-104, 118-138 involved in the formation of intra-chain disulfid bridges.
141

	Remarks

• RELIABILITY OF THE PROTEIN SEQUENCE: The group of type 2 cystatins consists of several proteins with highly similar sequence, including human cystatins S, SN, and SA, which are located in the cystatin multigene locus. The SAGE-tag could potentially characterize cystatin S, SN or a cystatin related to SA, so that no precise decision could be made on the type of cystatin the TEM14 tag stands for. The cystatin S protein sequence was analyzed exemplary.

• CLASSIFICATION: Type two cystatins (contain at least two characteristic interchain disulfid bonds). Type 2 cystatins include the human cystatins C, D, S, SN, and SA, which are all products of genes located in the cystatin multigene locus on chromosome 20.

• FUNCTION: Several hints indicate a role of cystein protease (Cathepsins S and K) and their inhibitors in matrix turnover (collagen I, III, elastin), next to metalloproteinases.
     The most abundant cystein protease inhibitor Cystatin C, has been implicated in vascular diseases. A direct link for cystatin S, SA and angiogenesis cannot be derived from previous observations. Indirect evidence can be derived from the expression of cystatin S, SA in tumor cells (ESTs, Parle-McDermott 2000) and from the implications of some of the known targets in cancer biology. Cathepsins B, H and L have been shown to participate in processes of tumor growth, vascularization, invasion and metastasis (Kos,2000).

• TARGETS: Known targets for Cystatin S are Papain, Ficin, Cathepsin C, but not Cathepsin B (which has been implicated in cancer biology).
  Known targets for Cystatin SA are papain, ficin, cathepsin C, cathepsin H, and cathepsin K, but not Cathepsin B.

	Sequence Information

TEM	TEM14
Reliability of Gen2Tag mapping	Cid->tag (Hs.56319 S) frequency: tem14_1: CATG-GTACACACAC=2/3(cDNAs) Cid->tag (Hs.245776 similar to SA) frequency: tem14_2: CATG-GTACACACAC=2/2 ESTs, Highly similar to CYTT_HUMAN CYSTATIN SA PRECURSOR [H.sapiens] Cid->tag (Hs.123114 SN) frequency: tem14_3: CATG-GTACACACAC=1/1 ESTs, cystatin SN REMARKS: Tem14_1 and Tem14_3 are clearly associated with known proteins and are discussed in the following. The information given through the tag is insufficient to decide on Tem14_1,_2 or _3.
Analysis	BlastP against nr Output Created by a Suite of Single Sequence Analysis Techniques
Protein Description	SEQ.SOURCE TEM14_1: sp|P01036|CYTS_HUMAN CYSTATIN S SEQ.SOURCE TEM14_3: sp|P01037|CYTN_HUMAN CYSTATIN SN REMARKS: outside the Signalpeptide the two sequences are highly similar (90% identity): thus the domainanalysis is performed exemplary on Cystatin S REMARKS: SIMILARITY: BELONG TO THE TYPE 2 CYSTATIN FAMILY. SIGNAL 1 20 PROBABLE. CHAIN 21 141 CYSTATIN SA. ACT_SITE 32 32 REACTIVE SITE. SITE 76 80 SECONDARY AREA OF CONTACT.
Protein Sequence	>TEM14_1_sp|P01036|CYTS_HUMAN CYSTATIN S PRECURSOR MARPLCTLLLLMATLAGALASSSKEENRIIPGGIYDADLNDEWVQRALHF AISEYNKATEDEYYRRPLQVLRAREQTFGGVNYFFDVEVGRTICTKSQPN LDTCAFHEQPELQKKQLCSFEIYEVPWEDRMSLVNSRCQEA >TEM14_3_sp|P01037|CYTN_HUMAN CYSTATIN SN PRECURSOR MAQHLSTLLLLLATLAVALAWSPKEEDRIIPGGIYNADLNDEWVQRALHF AISEYNKATKDDYYRRPLRVLRARQQTVGGVNYFFDVEVGRTICTKSQPN LDTCAFHEQPELQKKQLCSFEIYEVPWENRRSLVKSRCQES
SAGE-Tag	CATG-GTACACACACC
EST/cluster Description	MARKINGS: TEM14 Tag in red first Poly-A-signal following the Tag in bold
EST/cluster Sequence	TEM14_1: Hs.56319 CystatinS GGCTCTCACCCTCCTCTCCTGCAGCTCCAGCTCTGTGCTCTGCCTCTGAGGAGACCATGGCCCGGCCTCT GTGTACCCTGCTACTCCTGATGGCTACCCTGGCTGGGGCTCTGGCCTCGAGCTCCAAGGAGGAGAATAGG ATAATCCCAGGTGGCATCTATGATGCAGACCTCAATGATGAGTGGGTACAGCGTGCCCTTCACTTCGCCA TCAGCGAGTACAACAAGGCCACCGAAGATGAGTACTACAGACGCCCGCTGCAGGTGCTGCGAGCCAGGGA GCAGACCTTTGGGGGGGTGAATTACTTCTTCGACGTAGAGGTGGGCCGCACCATATGTACCAAGTCCCAG CCCAACTTGGACACCTGTGCCTTCCATGAACAGCCAGAACTGCAGAAGAAACAGTTATGCTCTTTCGAGA TCTACGAAGTTCCCTGGGAGGACAGAATGTCCCTGGTGAATTCCAGGTGTCAAGAAGCCTAGGGGTCTGT GCCAGGCCAGTCACACCGACCACCACCCACTCCCACCCCCTGTAGTGCTCCCACCCCTGGACTGGTGGCC CCCACCCTGCGGGAGGCCTCCCCATGTGCCTGTGCCAAGAGACAGACAGAGAAGGCTGCAGGAGTCCTTT GTTGCTCAGCAGGGCGCTCTGCCCTCCCTCCTTCCTTCTTGCTTCTAATAGACCTGGTACATGGTACACA CACCCCCACCTCCTGCAATTAAACAGTAGCATCGCC TEM14_2: Hs.245776.1: similar to Cystatin SA GGTGGCATCTATGATGCAGACCCAATAATAACGGGGTCAGCGTGCCCCTTCACTTTGTCATCAGCGAGTA TACAAGGGCCACTGAAGATGAGTACTACAGACGCTTCTGCGGGGTGCTACGAGCCAGGGAGCAGATCGTG GTCGGGGTGAATTACTTCTTCGACATAGAGGTGGGCCGAACCATATGTACCAAGTCNCAGCCCAACTTGG ACACCTGTGCCTTCCATGAACAGCCAGAACTGCAGAAGAAACAGTTGTGCTCTTTCCAGATCTACGAAGT TCCCTGGGAGGACAGAATGTCCCTGGTGAATTCCAGGTGTCAAGAAGCCTAGGGATCTGTGCCAGGGAGT CACACTGACCACCTCCTATTCCTACCCTTTGTAGTGTTCCCACCCTTGGATTGGTGGCCCCCACCTTGTG GGAGGTCTCCCCATGCACTTGCAGCAGGAGAAGACAGAGAAGGCTGCAGGAGGCCTTTGTTGCTCAGCAG GGGACTCTGCCCTCCCTCCTTCCTTTTGCTTCTCATAGCCCTGGTACATGGTACACACACCCCCACCTCC TGCAATTAAACAGTAGCATCACCTC TEM14_3: THC482794 CCCACCCTGGGACAGGCTCTCACCCTCCTCTCCTGCAGCTCCAGCTTTGTGCTCTGCCTCTGAGGAGACC ATGGCCCAGCATCTGAGTACCCTGCTGCTCCTGCTGGCCACCCTAGCTGTGGCCCTGGCCTGGAGCCCCA AGGAGGAGGATAGGATAATCCCGGGTGGCATCTATAACGCAGACCTCAATGATGAGTGGGTACAGCGTGC CCTTCACTTCGCCATCAGCGAGTATAACAAGGCCACCAAAGATGACTACTACAGACGTCCGCTGCGGGTA CTAAGAGCCAGGCAACAGACCGTTGGGGGGGTGAATTACTTCTTCGACGTAGAGGTGGGCCGAACCATAT GTACCAAGTCCCAGCCCAACTTGGACACCTGTGCCTTCCATGAACAGCCAGAACTGCAGAAGAAACAGTT GTGCTCTTTCGAGATCTACGAAGTTCCCTGGGAGAACAGAAGGTCCCTGGTGAAATCCAGGTGTCAAGAA TCCTAGGGATCTGTGCCAGGCCATTCGCACCAGCCACCACCCACTCCCACCCCCTGTAGTGCTCCCACCC CTGGACTGGTGGCCCCCACCCTGCGGGAGGCCTCCCCATGTGCCTGCGCCAAGAGACAGACAGAGAAGGC TGCAGGAGTCCTTTGTTGCTCAGCAGGGCGCTCTGCCCTCCCTCCTTCCTTCTTGCTTCTAATAGCCCTG GTACATGGTACACACACCCCCACCTCCTGCAATTAAACAGTAGCATCGCCGGGGAAAAGAATTAAACT
Genomic Sequence Description	SEQ.SOURCE: gi|9187269|emb|AL359433.7|AL359433 RP1-234M6
Genomic Sequence

	Literature

PubMed: 10952775	Serial analysis of gene expression identifies putative metastasis-associated transcripts in colon tumour cell lines. le-McDermott A, McWilliam P, Tighe O, Dunican D, Croke DT. Br J Cancer. 2000 Sep;83(6):725-8.
PubMed: 10545518	Cystatin C deficiency in human atherosclerosis and aortic aneurysms. Shi GP, Sukhova GK, Grubb A, Ducharme A, Rhode LH, Lee RT, Ridker PM, Libby P, Chapman HA. J Clin Invest. 1999 Nov;104(9):1191-7.
PubMed: 9691094	Expression of the elastolytic cathepsins S and K in human atheroma and regulation of their production in smooth muscle cells. Sukhova GK, Shi GP, Simon DI, Chapman HA, Libby P. J Clin Invest. 1998 Aug 1;102(3):576-83.
PubMed: 9587407	Production and characterization of two variants of human cystatin SA encoded by two alleles at the CST2 locus of the type 2 cystatin gene family. Saitoh E, Minaguchi K, Ishibashi O Arch Biochem Biophys 1998 Apr 15;352(2):199-206
PubMed: 10763147	Cysteine proteinases and their inhibitors in extracellular fluids: markers for diagnosis and prognosis in cancer. Kos J, Werle B, Lah T, Brunner N. Int J Biol Markers. 2000 Jan-Mar;15(1):84-9.