An environmental assessment of the John Pennekamp Coral Reef State Park and the Key Largo Coral Reef Marine Sanctuary (Unpublished 1983 Report)

The Pennekamp Coral Reef State Park was established in 1960 and the Key Largo National Marine Sanctuary in 1975. Field studies, funded by NOAA, were conducted in 1980 - 1981 to determine the state of the coral reefs and surrounding areas in relation to changing environmental conditions and resource management that had occurred over the intervening years. Ten reef sites within the Sanctuary and seven shallow grass and hardbottom sites within the Park were chosen for qualitative and quantitative studies. At each site, three parallel transects not less than 400 m long were run perpendicular to the reef or shore, each 300 m apart. Observations, data collecting and sampling were done by two teams of divers. Approximately 75 percent of the bottom within the 18-m isobath was covered by marine grasses, predominantly turtle grass. The general health of the seagrasses appeared good but a few areas showed signs of stress. The inner hardbottom of the Park was studied at the two entrances to Largo Sound. Though at the time of the study the North Channel hardbottom was subjected to only moderate boat traffic, marked changes had taken place over the past years, the most obvious of which was the loss of the extensive beds of Sargassum weed, one of the most extensive beds of this alga in the Keys. Only at this site was the green alga Enteromorpha encountered. This alga, often considered a pollution indicator, may denote the effects of shore run off. The hardbottom at South Channel and the surrounding grass beds showed signs of stress. This area bears the heaviest boat traffic within the Park waters causing continuous turbidity from boat wakes with resulting siltation. The offshore hardbottom and rubble areas in the Sanctuary appeared to be in good health and showed no visible indications of deterioration. Damage by boat groundings and anchors was negligible in the areas surveyed. The outer reefs in general appear to be healthy. Corals have a surprising resiliency to detrimental factors and, when conditions again become favorable, recover quickly from even severe damage. It is, therefore, a cause for concern that Grecian Rocks, which sits somewhat inshore of the outer reef line, has yet to recover from die-off in 1978. The slow recovery, if occurring, may be due to the lower quality of the inshore waters. The patch reefs, more adapted to inshore waters, do not show obvious stress signs, at least those surveyed in this study. It is apparent that water quality was changing in the keys. Water clarity over much of the reef tract was observed to be much reduced from former years and undoubtedly plays an important part in the stresses seen today over the Sanctuary and Park. (PDF contains 119 pages)

Olympic Coast National Marine Sanctuary Habitat Mapping: Survey report and classification of side scan sonar data from surveys HMPR-114-2004-02 and HMPR-116-2005-01.

The Olympic Coast National Marine Sanctuary (OCNMS) continues to invest significant resources into seafloor mapping activities along Washington’s outer coast (Intelmann and Cochrane 2006; Intelmann et al. 2006; Intelmann 2006). Results from these annual mapping efforts offer a snapshot of current ground conditions, help to guide research and management activities, and provide a baseline for assessing the impacts of various threats to important habitat. During the months of August 2004 and May and July 2005, we used side scan sonar to image several regions of the sea floor in the northern OCNMS, and the data were mosaicked at 1-meter pixel resolution. Video from a towed camera sled, bathymetry data, sedimentary samples and side scan sonar mapping were integrated to describe geological and biological aspects of habitat. Polygon features were created and attributed with a hierarchical deep-water marine benthic classification scheme (Greene et al. 1999). For three small areas that were mapped with both side scan sonar and multibeam echosounder, we made a comparison of output from the classified images indicating little difference in results between the two methods. With these considerations, backscatter derived from multibeam bathymetry is currently a costefficient and safe method for seabed imaging in the shallow (<30 meters) rocky waters of OCNMS. The image quality is sufficient for classification purposes, the associated depths provide further descriptive value and risks to gear are minimized. In shallow waters (<30 meters) which do not have a high incidence of dangerous rock pinnacles, a towed multi-beam side scan sonar could provide a better option for obtaining seafloor imagery due to the high rate of acquisition speed and high image quality, however the high probability of losing or damaging such a costly system when deployed as a towed configuration in the extremely rugose nearshore zones within OCNMS is a financially risky proposition. The development of newer technologies such as intereferometric multibeam systems and bathymetric side scan systems could also provide great potential for mapping these nearshore rocky areas as they allow for high speed data acquisition, produce precisely geo-referenced side scan imagery to bathymetry, and do not experience the angular depth dependency associated with multibeam echosounders allowing larger range scales to be used in shallower water. As such, further investigation of these systems is needed to assess their efficiency and utility in these environments compared to traditional side scan sonar and multibeam bathymetry. (PDF contains 43 pages.)

Olympic Coast National Marine Sanctuary Area to be Avoided (ATBA) Education and Monitoring Program

The National Marine Sanctuaries Act (16 U.S.C. 1431, as amended) gives the Secretary of Commerce the authority to designate discrete areas of the marine environment as National Marine Sanctuaries and provides the authority to promulgate regulations to provide for the conservation and management of these marine areas. The waters of the Outer Washington Coast were recognized for their high natural resource and human use values and placed on the National Marine Sanctuary Program Site Evaluation List in 1983. In 1988, Congress directed NOAA to designate the Olympic Coast National Marine Sanctuary (Pub. L. 100-627). The Sanctuary, designated in May 1994, worked with the U.S. Coast Guard to request the International Maritime Organization designate an Area to be Avoided (ATBA) on the Olympic Coast. The IMO defines an ATBA as "a routeing measure comprising an area within defined limits in which either navigation is particularly hazardous or it is exceptionally important to avoid casualties and which should be avoided by all ships, or certain classes of ships" (IMO, 1991). This ATBA was adopted in December 1994 by the Maritime Safety Committee of the IMO, “in order to reduce the risk of marine casualty and resulting pollution and damage to the environment of the Olympic Coast National Marine Sanctuary”, (IMO, 1994). The ATBA went into effect in June 1995 and advises operators of vessels carrying petroleum and/or hazardous materials to maintain a 25-mile buffer from the coast. Since that time, Olympic Coast National Marine Sanctuary (OCNMS) has created an education and monitoring program with the goal of ensuring the successful implementation of the ATBA. The Sanctuary enlisted the aid of the U.S. and Canadian coast guards, and the marine industry to educate mariners about the ATBA and to use existing radar data to monitor compliance. Sanctuary monitoring efforts have targeted education on tank vessels observed transiting the ATBA. OCNMS's monitoring efforts allow quantitative evaluation of this voluntary measure. Finally, the tools developed to monitor the ATBA are also used for the more general purpose of monitoring vessel traffic within the Sanctuary. While the Olympic Coast National Marine Sanctuary does not currently regulate vessel traffic, such regulations are within the scope of the Sanctuary’s Final Environmental Impact Statement/Management Plan. Sanctuary staff participate in ongoing maritime and environmental safety initiatives and continually seek opportunities to mitigate risks from marine shipping.(PDF contains 44 pages.)

Southern California Baseline Studies and Analysis: 1975-1978. Master Table of Contents

This is only the table of contents for a series of technical reports done from 1975-1978. The papers were done on contract for BLM by a number of universities and consulting firms such as Science Applications, Inc., University of Southern California, Scripps Institute of Oceanography, Moss Landing Marine Laboratories, and various campuses of University of California and California State University. (PDF contains 36 pages)

Spawning, egg development, and early life history dynamics of arrowtooth flounder (Atheresthes stomias) in the Gulf of Alaska

Arrowtooth flounder (Atheresthes stomias) has the highest biomass of any groundfish species in the Gulf of Alaska, is a voracious predator of age 1 walleye pollock (Theragra chalcogramma), and is a major component in the diet of Steller sea lions (Eumetopias jubatus). Owing to its ecological importance in the Gulf of Alaska and the limited information available on its reproduction, interest has intensified in describing its spawning and early life history. A study was undertaken in late January–February 2001–2003 in the Gulf of Alaska to obtain information on adult spawning location, depth distribution, and sexual maturity, and to obtain fertilized eggs for laboratory studies. Adults were found 200–600 m deep east of Kodiak Island over the outer continental shelf and upper slope, and southwest along the shelf break to the Shumagin Islands. Most ripe females (oocytes extruded with light pressure) were found at 400 m and most ripe males (milt extruded with light pressure) were found at depths ≥450 m. Eggs were fertilized and incubated in the laboratory at 3.0°, 4.5°, and 6.0°C. Eggs were reared to hatching, but larvae did not survive long enough to complete yolk absorption and develop pigment. Eggs were staged according to morphological hallmarks and incubation data were used to produce a stage duration table and a regression model to estimate egg age based on water temperature and developmental stage. Arrowtooth flounder eggs (1.58–1.98 mm in diameter) were collected in ichthyoplankton surveys along the continental shelf edge, primarily at depths ≥400 m. Early-stage eggs were found in tows that sampled to depths of ≥450 m. Larvae, which hatch between 3.9 and 4.8 mm standard length, increased in abundance with depth. Observations on arrowtooth flounder eggs and early-stage larvae were used to complete the description of the published partial developmental series.(PDF file contains 34 pages.)

Seasonal, horizontal, and vertical distribution of phytoplankton chlorophyll a in the Northeast U.S. Continental Shelf Ecosystem

The broad scale features in the horizontal, vertical, and seasonal distribution of phytoplankton chlorophyll a on the northeast U.S. continental shelf are described based on 57,088 measurements made during 78 oceanographic surveys from 1977 through 1988. Highest mean water column chlorophyll concentration (Chlw,) is usually observed in nearshore areas adjacent to the mouths of the estuaries in the Middle Atlantic Bight (MAB), over the shallow water on Georges Bank, and a small area sampled along the southeast edge of Nantucket Shoals. Lowest Chlw «0.125 ug l-1) is usually restricted to the most seaward stations sampled along the shelf-break and the central deep waters in the Gulf of Maine. There is at least a twofold seasonal variation in phytoplankton biomass in all areas, with highest phytoplankton concentrations (m3) and highest integrated standing stocks (m2) occurring during the winter-spring (WS) bloom, and the lowest during summer, when vertical density stratification is maximal. In most regions, a secondary phytoplankton biomass pulse is evident during convective destratification in fall, usually in October. Fall bloom in some areas of Georges Bank approaches the magnitude of the WS-bloom, but Georges Bank and Middle Atlantic Bight fall blooms are clearly subordinate to WS-blooms. Measurements of chlorophyll in two size-fractions of the phytoplankton, netplankton (>20 um) and nanoplankton «20 um), revealed that the smaller nanoplankton are responsible for most of the phytoplankton biomass on the northeast U.S. shelf. Netplankton tend to be more abundant in nearshore areas of the MAB and shallow water on Georges Bank, where chlorophyll a is usually high; nanoplankton dominate deeper water at the shelf-break and deep water in the Gulf of Maine, where Chlw is usually low. As a general rule, the percent of phytoplankton in the netplankton size-fraction increases with increasing depth below surface and decreases proceeding offshore. There are distinct seasonal and regional patterns in the vertical distribution of chlorophyll a and percent netplankton, as revealed in composite vertical profiles of chlorophyll a constructed for 11 layers of the water column. Subsurface chlorophyll a maxima are ubiquitous during summer in stratified water. Chlorophyll a in the subsurface maximum layer is generally 2-8 times the concentration in the overlying and underlying water and approaches 50 to 75% of the levels observed in surface water during WS-bloom. The distribution of the ratio of the subsurface maximum chlorophyll a to surface chlorophyll a (SSR) during summer parallels the shelfwide pattern for stability, indexed as the difference in density (sigma-t) between 40 m and surface (stability 40. The weakest stability and lowest SSR's are found in shallow tidally-mixed water on Georges Bank; the greatest stability and highest SSR's (8-12:1) are along the mid and outer MAB shelf, over the winter residual water known as the "cold band." On Georges Bank, the distribution of SSR and the stability40 are roughly congruent with the pattern for maximum surface tidal current velocity, with values above 50 cms-1 defining SSR's less than 2:1 and the well-mixed area. Physical factors (bathymetry, vertical mixing by strong tidal currents, and seasonal and regional differences in the intensity and duration of vertical stratification) appear to explain much of the variability in phytoplankton chlorophyll a throughout this ecosystem. (PDF file contains 126 pages.)

Benthic Macrofauna of the New York Bight, 1979-89

The benthic macrofauna of the New York Bight has been monitored extensively, primarily to determine trends over space and time in biological effects of waste inputs. In the present study, from 44 to 48 stations were sampled each summer from 1980-1985. Data from other Bight benthic studies are included to· extend the temporal coverage from 1979 to 1989. Numbers of species and amphipods per sample, taken as relatively sensitive indicators of environmental stress, showed consistent spatial patterns. Lowest values were found in the Christiaensen Basin and other inshore areas, and numbers increased toward the outermost shelf and Hudson Shelf Valley stations. There were statistically significant decreases in species and amphipods at most stations from 1980 to 1985. (Preliminary data from a more recent study suggest numbers of species increased again between 1986 and 1989.) Cluster analysis of 1980-85 data indicated several distinct assemblages-sewage sludge dumpsite, sludge accumulation area, inner Shelf Valley, outer Shelf Valley, outer shelf-with little change over time. The "enriched" and "highly altered" assemblages in the Basin appear similar to those reported since sampling began there in 1968. No consistently defaunated areas have been found in any sampling programs over the past 20 years. On a gross level, therefore, recent faunal responses to any environmental changes are not evident, but the more sensitive measures used, i.e. numbers of species and amphipods, do indicate widespread recent effects. Causes of the faunal changes are not obvious; some possibilities, including increasing effects of sewage sludge or other waste inputs, natural factors, and sampling artifacts, are discussed. (PDF file contains 54 pages.)

Marine Flora and Fauna of the Northeastern United States: Erect Bryozoa

Forty-nine species of erect Bryozoa from a broad range of Cyclostome, Ctenostome, and Cheilostome families are described and illustrated, and an artificial dichotomous key is provided for their identification. In general, the marine bryozoan faunas of the northeastern coasts of the United States are poorly known; species records are sparse and voucher collections few, and it is certain that many more species occur in this region than are presently known. The species described here occur in intertidal, coastal or offshore habitats; some are well known and have been recorded on numerous previous occasions, others have been only rarely reported, while a few are known to occur commonly in the north of the region but have yet to be recorded south of Cape Cod. Some of the species described have not been recorded at all on northeastern coasts of the United States, but are widely distributed in North Atlantic continental shelf habitats and perhaps occur in similar parts of the outer shelf of this region. This fauna is thus provisional, but is intended to stimulate further work on the Bryozoa. (PDF file contains 52 pages.)

Early-life-history profiles, seasonal abundance, and distribution of four species of Carangid larvae off Louisiana, 1982 and 1983

We present data on ichthyoplankton distribution, abundance, and seasonality and supporting environmental information for four species of coastal pelagics from the family Carangidae: blue runner Caranx crysos, Atlantic bumper Chloroscombrus chrysurus, round scad Decapterus punctatus, and rough scad Trachurus lathami. Data are from 1982 and 1983 cruises off Louisiana sponsored by the Southeastern Area Monitoring and Assessment Program (SEAMAP). Bioprofiles on reproductive biology, early life history, meristics, adult distribution, and fisheries characteristics are also presented for these species. Maximum abundances of larval blue runner, Atlantic bumper, and round scad were found in July inside the 4O-m isobath, although during the rest of the cruises these species were rarely found together. Larval Atlantic bumper were captured in June and July only; blue runner in May, June, and July; and round scad in all seasons. Atlantic bumper larvae, concentrated mostly off western Louisiana, were by far the most abundant carangid in 1982 and 1983. Larval blue runner were the second most abundant summer-spawned carangid in 1982 and 1983, but their abundance and depth distribution varied considerably between years. Relative abundance of larval round scad off Louisiana was low, and they were captured only west of the Mississippi River delta, although they are reported to dominate carangid populations in the eastern Gulf of Mexico. Rough scad were primarily winter/spring and outer-shelf (40-182 m) spawners. They ranked third in overall abundance, but were the most abundant target carangid on the outer shelf. Ecological parameters such as surface salinity, temperature, and station depth are presented from capture sites for recently hatched larvae <2.5 mm notochord length, except round scad) as well as for all sizes of fish below 14 mm standard length. (PDF file contains 44 pages.)

Early-life-history profiles, seasonal abundance, and distribution of four species of Clupeid larvae from the northern Gulf of Mexico, 1982 and 1983

We present data on ichthyoplankton distribution, abundance, and seasonality and supporting environmental information for four species of coastal pelagics from the family Clupeidae: round herring Etrumeus teres, scaled sardine Harengula jaguana, Atlantic thread herring Opisthonema oglinum, and Spanish sardine Sardinella aurita. Data are from 1982 and 1983 cruises across the northern Gulf of Mexico sponsored by the Southeastern Area Monitoring and Assessment Program (SEAMAP). This is the first such examination for these species on a multiyear and gulfwide scale. Bioproflles on reproductive biology, early life history, meristics, adult distribution, and fisheries characteristics are also presented for these species. During the summer, larval Atlantic thread herring and scaled and Spanish sardines were abundant on the inner shelf <40 m depth), but were rare or absent in deeper waters. Scaled sardine and thread herring were found virtually everywhere inner-shelf waters were sampled, but Spanish sardines were rare in the north-central Gulf. During 1982, larval Atlantic thread herring were the most abundant of the four target c1upeid species, whereas Spanish sardine were the most abundant during 1983. On the west Florida shelf, Spanish sardine dominated larval c1upeid populations both years. Scaled sardine larvae were the least abundant of the four species both years, but were still captured in 25% of inner-shelf bongo net collections. Round herring larvae, collected February-early June (primarily March-April), were abundant on the outer shelf (40-182 m depth) and especially off Louisiana. Over the 2-year period, outer-shelf mean abundance for round herring was 40.2 larvae/10 m2; inner-shelf mean abundances for scaled sardine, Atlantic thread herring, and Spanish sardine were 14.9, 39.2, and 41.9 larvae/l0 m2, respectively. (PDF file contains 66 pages.)

Atlas and zoogeography of common fishes in the Bering Sea and northeastern Pacific

The geographic and depth frequency distribution of 124 common demersal fish species in the northeastern Pacific were plotted from data on me at the Northwest and Alaska Fisheries Center (NWAFC), National Marine Fisheries Service. The data included catch records of fishes and invertebrates from 24,881 samples taken from the Chukchi Sea, throughout the Bering Sea, Aleutian Basin, Aleutian Archipelago, and the Gulf of Alaska, and from southeastern Alaska south to southern California. Samples were collected by a number of agencies and institutions over a 30-year period (1953-83), but were primarily from NWAFC demersal trawls. The distributions of all species with 100 or more occurrences in the data set were plotted by computer. Distributions plotted from these data were then compared with geographic and depth-range limits given in the literature. These data provide new range extensions (geographic, depth, or both) for 114 species. Questionable extensions are noted, the depth ranges determined for 95% of occurrences, and depths of most frequent occurrence are recorded. Ranges of the species were classified zoogeographically, according to life zone, and with regard to the depth zone of greatest occurrence. Because most species examined have broad geographic ranges, they do not provide the best information for testing the validity of proposed zoogeographic province boundaries. Because of the location of greatest sampling effort and methods used in sampling, most fIShes examined were eastern boreal Pacific, sublittoral-bathyal (outer shelf) species. (PDF file contains 158 pages.)

History of whaling in and near North Carolina

This study aims to reconstruct the history of shore whaling in the southeastern United States, emphasizing statistics on the catch of right whales, Eubalaena glacialis, the preferred targets. The earliest record of whaling in North Carolina is of a proposed voyage from New York in 1667. Early settlers on the Outer Banks utilized whale strandings by trying out the blubber of carcasses that came ashore, and some whale oil was exported from the 1660s onward. New England whalemen whaled along the North Carolina coast during the 1720s, and possibly earlier. As some of the whalemen from the northern colonies moved to Nortb Carolina, a shore-based whale fishery developed. This activity apparently continued without interruption until the War of Independence in 1776, and continued or was reestablished after the war. The methods and techniques of the North Carolina shore whalers changed slowly: as late as the 1890s they used a drogue at the end of the harpoon line and refrained from staying fast to the harpooned whale, they seldom employed harpoon guns, and then only during the waning years of the fishery. The whaling season extended from late December to May, most successfully between February and May. Whalers believed they were intercepting whales migrating north along the coast. Although some whaling occurred as far north as Cape Hatteras, it centered on the outer coasts of Core, Shackleford, and Bogue banks, particularly near Cape Lookout. The capture of whales other than right whales was a rare event. The number of boat crews probably remained fairly stable during much of the 19th century, with some increase in effort in the late 1870s and early 1880s when numbers of boat crews reached 12 to 18. Then by the late 1880s and 1890s only about 6 crews were active. North Carolina whaling had become desultory by the early 1900s, and ended completely in 1917. Judging by export and tax records, some ocean-going vessels made good catches off this coast in about 1715-30, including an estimated 13 whales in 1719, 15 in one year during the early 1720s, 5-6 in a three-year period of the mid to late 1720s, 8 by one ship's crew in 1727, 17 by one group of whalers in 1728-29, and 8-9 by two boats working from Ocracoke prior to 1730. It is impossible to know how representative these fragmentary records are for the period as a whole. The Carolina coast declined in importance as a cruising ground for pelagic whalers by the 1740s or 1750s. Thereafter, shore whaling probably accounted for most of the (poorly documented) catch. Lifetime catches by individual whalemen on Shackleford Banks suggest that the average annual catch was at least one to two whales during 1830·80, perhaps about four during the late 1870s and early 1880s, and declining to about one by the late 1880s. Data are insufficient to estimate the hunting loss rate in the Outer Banks whale fishery. North Carolina is the only state south of New Jersey known to have had a long and well established shore whaling industry. Some whaling took place in Chesapeake Bay and along the coast of Virginia during the late 17th and early 18th centuries, but it is poorly documented. Most of the rigbt whales taken off South Carolina, Georgia, and northern Florida during the 19th century were killed by pelagic whalers. Florida is the only southeastern state with evidence of an aboriginal (pre-contact) whale fishery. Right whale calves may have been among the aboriginal whalers' principal targets. (PDF file contains 34 pages.)

A temporal and spatial study of invertebrate communities associated with hard-bottom habitats in the South Atlantic Bight

Species composition, biomass, density, and diversity of benthic invertebrates from six bard-bottom areas were evaluated. Seasonal collections using a dredge, trawl, and suction and grab samplers yielded 432, 525, and 845 taxa, respectively. Based on collections wltb the different gear types, species composition of invertebrates was found to change bathymetrically. Inner- and mlddle-shelf sites were more similar to each other in terms of invertebrate species composition than they were to outer-shelf sites, regardless of season. Sites on the inner and outer shelf were grouped according to latitude; however, results suggest that depth is apparently a more important determinant of invertebrate species composition than either season or latitude. Sponges generally dominated dredge and trawl collections in terms of biomass. Generally, cnidarians, bryozoans, and sponges dominated at sites In terms of number of taxa collected. The most abundant smaller macrofauna collected in suction and grab samples were polychaetes, amphipods, and mollusks. Densities of the numerically dominant species changed botb seasonally and bathymetrically, with very few of these species restricted to a specific bathymetrlc zone. The high diversity of invertebrates from hard-bottom sites is attributed to the large number of rare species. No consistent seasonal changes in diversity or number of species were noted for individual stations or depth zones. In addition, H and its components showed no definite patterns related to depth or latitude. However, more species were collected at middle-shelf sites than at inner- or outer-shelf sites, which may be related to more stable bottom temperature or greater habitat complexity in that area. (PDF file contains 110 pages.)

Net phytoplankton and zooplankton in the New York Bight, January 1976 to February 1978, with comments on the effects of wind, Gulf Stream eddies, and slope water intrusions

Results are given of monthly net phytoplankton and zooplankton sampling from a 10 m depth in shelf, slope, and Gulf Stream eddy water along a transect running southeastward from Ambrose Light, New York, in 1976, 1977, and early 1978. Plankton abundance and temperature at 10 m and sea surface salinity at each station are listed. The effects of atmospheric forcing and Gulf Stream eddies on plankton distribution and abundance arc discussed. The frequency of Gulf Stream eddy passage through the New York Bight corresponded with the frequency of tropical-subtropical net phytoplankton in the samples. Gulf Stream eddies injected tropical-subtropical zooplankton onto the shelf and removed shelfwater and its entrained zooplankton. Wind-induced offshore Ekman transport corresponded generally with the unusual timing of two net phytoplankton maxima. Midsummer net phytoplankton maxima were recorded following the passage of Hurricane Belle (August 1976) and a cold front (July 1977). Tropical-subtropical zooplankton which had been injected onto the outer shelf by Gulf Stream eddies were moved to the inner shelf by a wind-induced current moving up the Hudson Shelf Valley. (PDF file contains 47 pages.)

A Computational Module Assembled from Different Protease Family Motifs Identifies PI PLC from Bacillus cereus as a Putative Prolyl Peptidase with a Serine Protease Scaffold

Proteolytic enzymes have evolved several mechanisms to cleave peptide bonds. These distinct types have been systematically categorized in the MEROPS database. While a BLAST search on these proteases identifies homologous proteins, sequence alignment methods often fail to identify relationships arising from convergent evolution, exon shuffling, and modular reuse of catalytic units. We have previously established a computational method to detect functions in proteins based on the spatial and electrostatic properties of the catalytic residues (CLASP). CLASP identified a promiscuous serine protease scaffold in alkaline phosphatases (AP) and a scaffold recognizing a beta-lactam (imipenem) in a cold-active Vibrio AP. Subsequently, we defined a methodology to quantify promiscuous activities in a wide range of proteins. Here, we assemble a module which encapsulates the multifarious motifs used by protease families listed in the MEROPS database. Since APs and proteases are an integral component of outer membrane vesicles (OMV), we sought to query other OMV proteins, like phospholipase C (PLC), using this search module. Our analysis indicated that phosphoinositide-specific PLC from Bacillus cereus is a serine protease. This was validated by protease assays, mass spectrometry and by inhibition of the native phospholipase activity of PI-PLC by the well-known serine protease inhibitor AEBSF (IC50 = 0.018 mM). Edman degradation analysis linked the specificity of the protease activity to a proline in the amino terminal, suggesting that the PI-PLC is a prolyl peptidase. Thus, we propose a computational method of extending protein families based on the spatial and electrostatic congruence of active site residues.